Exosome-associated microRNA as a diagnostic marker

(54) Title: ASSOCIATED MICRORNA AND EXOSOMES AS A DIAGNOSTIC MARKER (51) Int. Cl .: G01N 33/53 (30) Unionist Priority: 05/05/2008 US 61 / 050,438, 05/05/2008 US 61/050, 43825/07/2007 US 60 / 951,812 (73) Holder (s): UNIVERSITY OF LOUISVILLE RESEARCH FOUDATION, INC.

(72) Inventor (s): TAYLOR, DOUGLAS D .; GERCEL-TAYLOR, CICEK (74) Attorney (s): CRUZEIRO NEWMARC PATENTES E MARCAS LTDA.

(86) International Application: PCT US2008071235 of 25/07/2008 (87) International Publication: WO

2009/015357 of 01/29/2009

I. Apply a serum to the Sepharose 2B gel filtration column

2. Collect fractions of empty volurne containing exosomes *

4. Hybridize the probe marked with

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3. Isolate the circulating RNA exosomes obtained from the tumor, synthesize the cDNA probe for hybridization ^ by microarray.

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DIAGNOSIS

DESCRIPTION

RELATED REQUESTS

This order claims priority for Orders

North American Provisional Nos. 60 / 951,812 filed on July 2007, and 61 / 050,438 filed on May 5, 2008, the full disclosures of which are hereby incorporated by reference.

TECHNICAL FIELD

The material currently disclosed refers to methods for the diagnosis and prognosis of cancer and adverse pregnancy outcomes. In particular, the material disclosed here refers to diagnostic and prognostic methods based on the determination of quantities of one or more microRNAs derived from exosomes correlated with cancer or with adverse results of pregnancies in a biological sample of an individual.

HISTORIC

The identification of cancer biomarkers suitable for the previous detection and diagnosis of cancer holds great promise for improving the clinical outcome of individuals. It is especially important for individuals who have some symptoms or no symptoms at all, or with tumors that are already relatively inaccessible to physical examination. Despite considerable efforts directed at prior detection, some reliable and cost-effective screening tests have been developed that can diagnose cancer in the primary stage.

MICRORNA ASSOCIATED WITH EXOSOMES AS A MARKER ζ

As an example, ovarian cancer remains the sixth most common cancer in women worldwide, causing approximately 125,000 deaths annually (Sankaranarayanan &amp; Ferlay, 2006). Many women with ovarian cancer are diagnosed at an advanced stage, with 75% diagnosed with extraovarian disease (Berek et al., 2003). In comparison with other cancers associated with women, 73% of endometrial cancers, 55% of breast cancers and 50% of cervical cancers are diagnosed in Stage I of the disease (Menon &amp;

Jacobs, 2000). Although the 5-year survival rate of survivors with Stage I ovarian cancer exceeds 90%, only 21% of patients with advanced stage ovarian cancer survive for us after the initial diagnosis (Berek et al., 2003). Since long-term survival has not changed significantly in the past 15 decades, the best prospect for longer ovarian cancer survival lies in early diagnosis (Menon

Jacobs, 2000) ...

The only biomarker currently approved for the detection of ovarian cancer is CA125, and its quantification by

ELISA has been the gold standard for the detection of ovarian cancer since its introduction in 1983. The evaluation of CA125 is commonly used to treat the disease, both in detecting the disease and in monitoring the recurrence of the disease; however, the use of CA125 is limited to the early stages of cancer detection (sensitivity 50-60%). The CA125 quantification is only approved and consistently approved in monitoring remission.

CA125 is neither sensitive nor specific for ovarian cancer detection again, as it is elevated in&gt; 50% of women with stage I disease, despite being elevated in more than 80% of ovarian cancer patients advanced stage. CA125 has poor specificity, which is demonstrated by its elevation in association with benign and malignant breast and colon disease, peritoneal irritants and 5 benign gynecological diseases, among others.

New strategies have been introduced that facilitate proteomic analysis by greatly simplifying the separation of the pre-analytical sample and coupling it with mass spectrometry (MS) in the research to discover the biomarker. The surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOFMS) type SELDI-TOF mass spectrometry has received much attention for its use in the resolution of proteins in biological test bodies by linking chip arrays biochemically distinct proteins. In one technology, four serum proteins are examined by ELISA, while another technology uses mass spectrometry of seven specific serum components or general peptide patterns in the patient's serum to define the presence of cancer. The SELDI-TOF-MS profiling has been used successfully to differentiate ovarian, breast, prostate and liver cancer from controls.

Serum SELDI-TOF-MS profiling has been shown to be significantly better than the current standard CA125 serum biomarker in different patients with ovarian cancer than those with benign ovarian disease and healthy controls. The 25 studies demonstrated that selecting a multiple protein combination solved by SELDI-TOF-MS may have the potential for a diagnostic approach. An effective screening test for ovarian cancer should achieve high sensitivity and specificity and ι

currently, different proteomic technologies, as well as computer analytical tools used to discern peaks generated from different findings. These initial studies on the SELDI-TOF-MS profiling are promising, and the concept is reproducible in a number of different backgrounds; however, translating this approach into routine diagnostic tests remains difficult.

It has been calculated that in order to be an effective screening test, an assay must achieve a minimum of 99.6% specificity. To obtain this level of specificity, several components of the tumor characteristics must be incorporated without new diagnostic tests for effective detection, due to the multifactorial nature of ovarian cancer, as well as other cancers. A disadvantage of mass spectrometry techniques 15 is that some samples of importance can be masked by more abundant proteins in DM, as well as in the analysis of spectrometric production. Pre-purification by various techniques such as high performance liquid chromatography and positive or negative affinity binding can remove some 20 specific groups of proteins. The biggest challenge in most current mass spectrometry approaches is dynamic range rather than sensitivity. Although the removal of prevalent proteins or peptides can greatly increase the amount of information that can be obtained from certain samples, important proteins such as albumin can function as carriers of protein subarrays of diagnostic significance. Further studies with larger samples and careful blind mode of independent validation sets are required before any consideration of the application of this platform for screening ovarian cancer or any other indication is considered.

Thus, there remains a need to develop better biomarkers in almost all cancers and other disorders, including the increased risk of adverse pregnancy outcomes. Blood tests remain an attractive goal, due to the availability and ease of sample collection. Early defensive diagnoses of cancer and increased risk of adverse pregnancy outcomes would facilitate early and potentially more effective treatment of patients. Thus, there is a lack of biomarkers that can, individually in combination or with other biomarkers or diagnostic modalities, provide the necessary sensitivity and specificity 15 for the early detection and prognosis of cancer and adverse pregnancy results. In particular, simple examinations of cancer biomarkers and adverse pregnancy outcomes in rapidly accessible biological fluids are needed.

SUMMARY

This Summary lists several configurations of the material disclosed herein and in many cases may list variations and permutations of those configurations. This Summary is just an example of the numerous and various configurations. The mention of one or more characteristics representative of a particular configuration is likewise exemplary. This configuration can typically exist with or without the mentioned feature (s); also, these characteristics may apply to other configurations of the matter disclosed herein, whether or not they are listed in this Summary. To avoid excessive repetition, this Summary does not list or suggest all possible combinations of such characteristics.

In some configurations of the material now disclosed, a method is provided for the diagnosis of cancer in an individual.

In some configurations, the method comprises providing a biological sample from an individual; the isolation of cancer-derived exosomes comprising microRNAs (miRNAs) from the biological sample; determining an amount of one or more of the miRNAs; and comparing the amount of one or more miRNAs with one or more levels of miRNA control. 0 The individual is then diagnosed as having cancer if there is a measurable difference in the amount of one or more miRNAs from the cancer-derived exosomes when compared to one or more levels of miRNA control. In some configurations, the method further comprises selecting a treatment or modifying a treatment for cancer based on the amount of one or more determined miRNAs.

In other configurations of the subject now disclosed, a method is provided for assessing the effectiveness and / or progression of cancer treatment in an individual. In some configurations, the method comprises providing a series of biological samples from an individual over a period of time; the isolation of cancer-derived exosomes comprising miRNAs from the series of biological samples; determining an amount of one or more of the miRNAs in each of the biological samples in the series; and the determination of any measurable action in the amounts of one or more miRNAs in each of the biological samples in the series, in order to evaluate the effectiveness and / or progression of cancer treatment in the individual.

In yet other configurations of the matter now disclosed, a method is provided for the characterization of cancer in an individual. In some configurations, the method comprises providing a biological sample from an individual; the isolation of cancer-derived exosomes comprising miRNAs from the biological sample; determining an amount of one or more of the miRNAs; and comparing the amount of one or more miRNAs with one or more levels of miRNA control. Cancer is then characterized based on the measurable difference in the amount of one or more miRNAs from the cancer-derived exosomes when compared to the one or more levels of miRNA control. In some configurations, the characterization of cancer comprises the determination of a type, class and / or stage of the cancer. In addition, in some configurations, determining the amount of one or more miRNAs comprises determining the total amount of miRNA in cancer-derived exosomes.

In some configurations of these methods, cancer is a cancer selected from the group consisting of ovarian cancer, cervical cancer, breast cancer, endometrial cancer, colon cancer, prostate cancer, lung cancer, melanoma and pancreatic cancer.

In addition, in some of these methods, the isolation of cancer-derived exosomes still includes the use of size exclusion chromatography to isolate cancer-derived exosomes. In some configurations, the isolation of cancer-derived exosomes comprises the centrifugation of a chromatographic fraction comprising cancer-derived exosomes. The chromatographic fraction may in some configurations be an empty volume fraction.

Still in some configurations, cancer-derived exosomes are separated from non-cancer exosomes by immunoabsorbent capture using an anti-cancer antigen antibody, such as an anti-epithelial cell adhesion molecule (anti-EpCAM).

In some of these methods, determining the amount of one or more miRNAs involves labeling one or more miRNAs, and in some configurations, then capture the one or more miRNAs with one or more polynucleotide probes that selectively bind one or more miRNAs. In other configurations of these methods, determining the amount of one or more miRNAs involves using a real-time polymerase chain reaction to quantify the amount of one or more miRNAs. In addition, in some configurations of these methods, miRNAs are one or more miRNAs shown in Table 2 including, for example, one or more miRNAs selected from the group consisting of miR-21, miR-141, miR200a, miR-200b, miR -200c, miR-203, miR-205 and miR-214.

In yet other configurations of the subject now disclosed, a method is provided for the diagnosis of adverse pregnancy outcomes in an individual. In some configurations, the method comprises providing a biological sample from an individual; the isolation of exosomes comprising miRNAs from the biological sample; determining an amount of one or more of the miRNAs; and comparing the amount of one or more miRNAs with one or more levels of miRNA control. The individual is diagnosed with the result of adverse pregnancy if there is a measurable difference in the amount of one or more miRNAs from the exosomes when compared to one or more levels of miRNA control. In some settings, the result of adverse pregnancy is a disorder selected from the group consisting of premature rupture of membranes, pre-eclampsia, premature birth, restriction of intrauterine growth and recurrent loss of pregnancy.

In some of the configurations presented here, the individual is human. In addition, in some of the configurations disclosed herein, the biological sample comprises milk, blood, serum, plasma, ascites, cystic fluid, pleural fluid, peritoneal fluid, cerebrospinal fluid, tears, urine, saliva, phlegm or combinations thereof.

It is also an objective of the matter now revealed to use the miRNAs associated with exosomes as diagnostic markers. This objective is achieved in whole or in part by the matter now revealed.

Having stated above an objective of the material disclosed in the present, and which is achieved in whole or in part by the material disclosed in the present, other objectives and advantages will become evident to the technicians in the subject after a study of the following description of the material, figures and the non-limiting examples disclosed herein.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 is a schematic diagram showing the exemplary method of chromatographic isolation of exosomes derived from cancer and miRNA from exosomes, determining amounts of miRNA by microarray technique, and analyzing the data to determine whether cancer is present in the examined individual.

Figure 2 is a schematic diagram showing the exemplary methodology for isolating exosomes derived from cancer and miRNA from exosomes, determining amounts of miRNA by PCR in real time, and analyzing the data to determine whether the cancer is present and the stage of the cancer in the examined individual.

Figure 3A is a graph showing the levels of the circulating exosomes derived from the tumor compared to the stage of ovarian cancer. Exosomes were isolated from sera obtained from female controls of combined ages (n = 10), women of the same age with benign ovarian disease (n = 10), and women diagnosed with ovarian cancer (n = 10 for each stage). The levels of exosomes presented as protein concentrations.

Figure 3B is an electronic micrograph of the circulating exosomes isolated by magnetic beads. Ultrafine sections (65 nm) were cut and colored with uranyl acetate and Reynold's lead citrate. The sections were examined using a Jeol 1210 transmission electron microscope.

Figure 4A is a graph showing the presence of small RNAs associated with positive EpCAM circulating exosomes from patients with ovarian cancer. A representative analysis of the RNA isolated from the tumor exosomes is shown using the

Agilent 2100 Bioanalyzer.

Figure 4B is a photograph of a separation of agarose gel (1%) of total RNA from the circulating exosomes and corresponding tumors. This total RNA was used as starting material for the miRNA profiling.

Figure 5 is a series of graphs showing the intensities of specific miRNAs obtained from advanced stage ovarian tumors (□) and EpCAM positive exosomes () isolated from the serum of these same patients. miR-21, miR-141, miR200a, miR-200b, miR-200c, miR-203, miR-205, miR-214 have been shown to be regulated upward markers of ovarian cancer. Each bar presents the average intensities of samples in duplicate with the results of four representative patients presented.

Figure 6 is a graph showing the intensities of specific miRNAs obtained from EpCAM-positive exosomes isolated from peripheral blood (2.5 mL) from patients with benign ovarian disease and from patients with ovarian cancer. Patients with ovarian cancer were separated between Stages

I, II and III. The bars represent the standard + mean deviation of the normalized intensities for each group of patients (n = 10 for each group).

Figures 7A and 7B are graphs showing a comparison of the specific exosomal miRNAs obtained from the serum of a patient with ovarian cancer, immediately after blood collection or 24, 48 and 96 hours after serum storage at 4 ° C (Figure 7A ) or after 7 to 28 days, stored at -70 ° C (Figure 7B). The tumor exosomes were isolated by MACS using antiEpCAM.

Figure 8 is a schematic diagram showing the exemplary methodology for the isolation of exosomes derived from cancer and miRNA from exosomes, determining amounts of miRNA by microarray technique, and analyzing the data to determine whether there is cancer in the examined individual.

Figure 9 is a series of graphs showing the intensities of specific miRNAs obtained from advanced lung tumors (light gray) and EpCAM-positive exosomes (dark gray) isolated from the serum of these same patients. Each bar presents the average intensities of samples in duplicate with the results of four representative patients presented.

DETAILED DESCRIPTION

Details of one or more configurations of the material disclosed herein are presented in the accompanying description below. Other characteristics, objectives and advantages of the material disclosed herein will be apparent from the specification, Figures and Claims. All publications, patent applications, patents and other references noted herein are incorporated by reference in their entirety. In case of conflict, you will have control over this specification, including definitions.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as common understanding by technicians on the subject to which the matter disclosed herein belongs. Although any methods, devices and materials similar or equivalent to those described in practice or in the testing of the material disclosed herein may be used, representative methods and materials will now be described.

In keeping with the enduring patent law convention, the terms one, one, and / or refer to one or more when used in this application, including the claims.

Thus, for example, the reference to a peptide includes a plurality of such peptides, and so on.

Unless otherwise stated, all numbers that express quantities of principles, conditions of reactions and others used in the specification and claims are to be understood as being modified in all examples by the term about. Thus, unless otherwise indicated, the numerical parameters indicated in this specification and in the appended claims are approximations that may vary depending on the properties desired and sought by the matter disclosed herein.

As used herein, the term about, when referring to a value or amount of mass, weight, time, volume, concentration or percentage means to encompass variations, in some configurations, of ± 20%, in some configurations of ± 10%, in some settings of ± 5%, in some settings of ± 1%, in some settings of ± 0.5% and, in some settings of ± 0.1% of the specified quantity, as these variations are suitable for carrying out the revealed methods.

In the past 5 years, expression profiling technology has identified new biomarkers with diagnostic applications. One of these biomarker groups is a class of small non-coding RNAs, called microRNAs (miRNAs) (Lorio et al. 2007; De Cecco et al., 2004; Calin &amp; Croce, 2006).

The microRNAs, small (22-25 nucleotides in length) non-coding RNAs, suppress the translation of the target mRNAs by binding to their 3 'untranslated regions (Esquela-Kerscher &amp; Slack, 2006;

Bartel, 2004). Post-transcriptional silencing of target genes by miRNA can occur either by dividing the homologous mRNA or by specific inhibition of protein synthesis.

All tumors analyzed by miRNA profiling showed significantly different miRNA signatures, compared to normal cells in the same tissue (Lorio et al.

2007; Calin &amp; Croce, 2006a; Calin &amp; Croce, 2006b). Lu et al.

(2005) performed an analysis of leukemias and solid cancers and determined that miRNA expression profiles could classify human cancers by developmental lineage and state of differentiation. Expressions of the signatures of individual miRNAs and specific miRNAs would now be linked to the diagnosis and prognosis of many human cancers.

Using tissue samples, lorio et al.

(2007) demonstrated that, compared to 'the normal ovary, specific miRNAs were expressed in an aberrant way in ovarian cancer, with miR-141, miR-200a, miR-200b and miR-200c being excessively more significantly expressed . They also demonstrated hypomethylation in ovarian tumors as a result of upward modulation of miR-21, miR-203 and miR-205, compared to the normal ovary. Two of these miRNAs, miR-200a and miR-200c modulated upwards were highlighted in all three histological types examined (serous, endometrioid and clear cells), considering that the upward modulation of miR-200b and miR-141 was shared by endometrioid and serous histological types. In general, miRNA signatures obtained by comparing the different histological types of ovarian cancers (serous, endometrioid, clear cells and mixed) with normal tissue were overlapped in most cases. Their analysis of ovarian tumors also demonstrated the absence of miRNAs differentially expressed with respect to the tumor stage or class, which could have resulted from their sample sets being obtained primarily from tumors in advanced stages.

Among the most significantly up-regulated miRNAs, miR-200a and miR-141 belong to the same family, miR-200b is located on chromosome 1 p36.33 in the same region as miR-200a and miR-200c is located on chromosome 12pl3.31 in the same region as miR-141 (Lorio et al. (2007)). This association would be in accordance with the findings of Zhang et al.

(2006) who proposed that upward modulation of specific miRNAs could be the amplification of miRNA genes. Using array-based high-resolution genomic hybridization, an aberrantly high proportion of loci containing miRNA genes exhibited changes in the number of DNA copies. In ovarian cancer, 37.1% of the genomic loci containing miRNA genes were associated with changes in the number of DNA copies (Zhang et al., 2006). In breast cancer and melanoma, an even greater proportion of these loci exhibit altered DNA copy numbers (72.8% and 85.9%, respectively) (Zhang et al., 2006). As a result, miRNA expression patterns or signatures appear to be more characteristic of the origins of tumor development than miRNA expression patterns and may be associated with diagnosis, staging, progression, prognosis and response to treatment.

However, as cancer diagnostic tools, before the material revealed in the present, the analysis of miRNA signatures was limited to tissue biopsies.

A recently described characteristic of cancer cells is the ability to release, or from intact shed, vesicular portions of the plasma membrane (referred to in the present exosomes, and also known in the art as fragments of membranes, membranous vesicles or microvesicules). Data that reveal, surprisingly for the first time, the miRNAs associated with exosomes originating from cancer cells (that is, exosomes derived from cancer) are revealed in the present study. The material revealed in the present still reveals for the first time that the miRNA isolated from cancer-derived exosomes demonstrates levels of expression in individuals suffering from cancer and that differ from the levels of miRNA expression (for example, increased or decreased) measured in individuals free from cancer (currently referred to as miRNA control levels). In addition, the material disclosed in the present provides the isolation of cancer-derived exosomes from the biological fluids readily accessible to an examined individual.

Thus, the material disclosed in the present provides, for the first time, methods for the diagnosis and prognosis of cancer based on the collection and measurement of exosomal miRNA levels derived from cancer from biological samples readily accessible, without the need for direct sampling of cancer cells. cancer.

Exosomes are microvesicles released from a variety of different cells, including cancer cells (that is, cancer-derived exosomes). These small vesicles (50 - 100nm in diameter) come from larger multivesicular endosomes and are secreted in the extracellular environment. The exact mechanism of exosome release / shedding remains unclear; however, this release is a phenomenon that requires energy, modulated by extracellular signals. They seem to be formed by the invagination and sprouting of the limiting membrane of the last endosomes, resulting in vesicles that contain cytosol and that expose the extracellular domain of cell proteins bound by membranes on its surface. Using electron microscopy, studies have shown the fusion profiles of multivessel endosomes with the plasma membrane, leading to the secretion of internal vesicles in the extracellular environment. The rate of exosome release is significantly increased in most neoplastic cells and occurs continuously. The increasing release of exosomes and their accumulation seem to be important in the process of malignant transformation. In addition to cancer cells, exosome release has also been shown to be associated with cells of embryonic origin (such as piacenta) and activated lymphoid cells.

Although extracellular shedding of exosomes occurs in other types of cells, under specific physiological conditions, the accumulation of exosomes from neoneoplastic cells is rarely observed in vivo. In contrast, the exosomes released by tumor cells accumulate in biological fluids, including serum, ascites and pleural fluids. The released exosomes and their accumulation seem to be important features of the malignant transformation. Cancer-derived shed exosomes do not reflect the overall composition of the plasma membrane of the tumor cell of origin, but represent micromaps with greater expression of tumor antigens.

The release of exosomes seems to be an important feature of intercellular communication. As the released exosomes express molecules with biological activity (such as Fas ligand, PD-1, MICA / B, mdrl, MMPs, CD44 and autoreactive antigens), the ability of these microvesicles to modulate lymphocyte and monocyte functions was analyzed in several models.

It has been theorized that these released exosomes modulate lymphocyte functions by mimicking activation-induced cell death (AICD).

Lymphoid cells appear to release exosomes after activation and appear to play an essential role in immunoregulation, preventing excessive immune responses and the development of autoimmunity. It has been postulated that the release of exosomes by tumor cells is a reexpression of fetal cell exosomes and that both constitute pathways for the surrounding immunovigilance.

MicroRNAs are small, naturally occurring non-coding RNAs that have lengths of about 17 to about 25 nucleotide bases (nt) in their biologically active forms. MiRNAs post-transcriptionally regulate gene expression by repressing the translation of the target mRNA. It is believed that the function of miRNAs as negative regulators, that is, greater amounts of a specific miRNA will be correlated with lower levels of expression of the target gene.

There are three forms of miRNAs existing in vivo, primary miRNAs (pri-miRNAs), premature miRNAs (pre-miRNAs) and mature miRNAs. Primary miRNAs (pri-miRNAs) are expressed as structured transcriptions in a stem-loop of about a few hundred bases up to 1 kb. The pri-miRNA transcripts are cleaved in the nucleus by an RNase II endonuclease called

Drosha, which cleaves both strands of the stem near the base of the stemloop. Drosha cleaves the duplex RNA with staggered cuts, leaving a 5 'phosphate and 2 nt pendant at the 3' end. The divination product, premature miRNA (premiRNA) is about 60 to about 110 nt long with a clamp structure formed folded back. The pre-miRNA is transported from the nucleus to the cytoplasm by

Ran-GTP and Exportin-5. Pre-miRNAs are further processed in the cytoplasm by another RNase II endonuclease called Dicer. THE

Dicer recognizes the 5 'phosphate and 3' pendant, and cleaves the loop to the outside of the stem-loop bond and forms duplex miRNA. The miRNA duplex binds to the RNA-induced silencing complex (RISC), where the antisense strand is preferably degraded and the mature sense stranded miRNA directs the RISC to its target site. Mature miRNA is the biologically active form of miRNA being about 17 to about nt long.

MicroRNAs work by coupling base pairs (perfect or imperfect) to specific sequences in their target gene messages (mRNA). MiRNA degrades or represses mRNA translation, causing the expression of target genes to be post-transcriptionally down-regulated, repressed or silenced. In animals, miRNAs do not necessarily have perfect homologies with their target sites, and partial homologies lead to translational repression, whereas in plants, where miRNAs tend to show complete homologies to their target sites, message degradation (mRNA) prevails. .

MicroRNAs are widely distributed in the genome, dominate genetic regulation and actively participate in many physiological and pathological processes. For example, it was determined that the regulatory modality of certain miRNAs controls cell proliferation, differentiation and apoptosis; and that abnormal miRNA profiles are associated with oncogenesis. In addition, it has been suggested that viral infection causes an increase in miRNAs designed to silence pro-cellular survival genes, and a reduction in miRNAs that suppress genes associated with apoptosis (programmed cell death), thus imbalancing in the direction of obtaining of apoptosis signaling.

Thousands of mRNA are under this selection pressure by hundreds of miRNA species identified so far. This selection process is instrumental in dampening specific groups of genetic expressions that, for example, may no longer be necessary, to allow cells to channel their directions from physiological programs to a new path of gene expression. The miRNA buffering dependent on gene expression target groups is a robust and fast regulation to allow cells to move from an old program and transition to a new one. A typical example of this is demonstrated during embryonic development, when a particular group of cells is directed to become specialized specialized cell types such as neurons, cardiomyocytes, muscles, etc.

It is believed that the expression levels of approximately one third of human genes are regulated by miRNAs, and that the miRNA regulation of exclusive gene expressions is linked to the particular signaling pathway for each specific cell type. For example, the apoptosis signaling pathway can be dictated by a group of miRNAs aimed at destabilizing pro-survival genetic messages, allowing alternative pro-apoptosis genes to dominate and thus activate the death program. Another example is the control of cancer growth; a recent finding showed that miRNAs can also be essential to prevent cells from becoming neoplastic. For example, two oncogenes, cMyc and cRas, were found to share control of a miRNA species, whose expression is down-regulated in cancer. In other words, the lack of this miRNA allows unchecked expression of cMyc and cRas, thus allowing these two genes to become abundantly present in cancer cells, allowing them to acquire an uncontrolled cell proliferation capacity and trigger the growth stage neoplastic. In addition, it has been reported that a miRNA mutation is responsible for a muscle phenotype in sheep of Belgian origin, suggesting that mutations associated with genetic disorders can be found in miRNAs, where no evidence of mutations in promoter regions, coding areas has been found. and slicing sites.

It is possible that a coordinated multipath orchestration serves to control a given cellular state, characterized by the fact that certain molecular hubs may be involved, which are functionally manipulated by hierarchical orders and redundancy of molecular control. In fact, dozens of miRNAs can operate to ensure that these hubs can perform large or small functions in cells, simply by suppressing the expression of themselves or their functional opponents. Thus, a genetic product can function as a main hub for a signaling path in one type of cell, and in another type of cell it can be a secondary hub, or it may not even be used. 0 microRNA control of the genetic expressions of the hub can then be an expedient mechanism to provide this versatility so that various molecules serve as both primary and secondary hubs, or in no way for different types of cellular operating modalities.

Given the role of miRNAs in genetic regulation, and in many physiological and pathological processes, it is desirable to obtain information about their interactive modes and patterns of expression. The systems and methods for quantifying and identifying which groups of alleged miRNAs are in operation in a given cell type, or in association with a particular process or condition of interest, can provide useful information for understanding how each cell state evolves and is maintained and how dysfunctional maintenance is prompted by inadequate reductions or increases in unique miRNA arrays 15 for the regulation of expression of major genes. This understanding can prove to be useful in the diagnosis and characterization of various disorders, including cancer and adverse pregnancy outcomes ...

As potential clinical diagnostic tools, miRNAs have proven to be important and accurate determinants for many, if not all cancers.

Increasing evidence shows that the expression of miRNA genes is deregulated in human cancer. The expression of miRNAs is highly specific for tissues and developmental stages, having recently allowed the molecular classification of tumors. To date, all tumors analyzed by miRNA profiling have demonstrated significantly different miRNA profiles when compared to normal cells in the same tissue. The profiling of miRNA by flow cytometry demonstrated that the profiles of miRNA expression classify human cancers according to the developmental lineage and the differentiation status of the tumors.

Excessive expression or low specific expression has been shown to correlate with certain types of tumors. Excessive microRNA expression can result in down regulation of tumor suppressor genes, considering that their underexpressions could lead to up regulation of the oncogene. Using large-scale microarray analysis, cancer cells showed distinct miRNA profiles when compared to normal cells with 36 of the 228 overexpressed miRNA genes and 21 down regulated in cancer cells versus normal cells.

Hierarchical cluster analyzes showed that this miRNA signature allowed tumor samples to be grouped based on their tissues of origin. Genomic scanning profiling studies have been carried out on various types of cancer, including CLL, breast cancer, glioblastoma, thyroid papillary carcinoma, hepatocellular carcinoma, ovarian cancer, colon cancer and endocrine pancreatic tumors. In a study of 104 combined pairs of cancerous and non-cancerous ovarian tissue, 43 differentially expressed miRNAs were observed; 28 were down regulated and 15 were overexpressed in tumors.

Statistical analyzes of microarray data obtained by two different methods, analysis of microarray significance (SAM) and analysis of prediction of microarray (MAP) of six solid tumors (ovarian, breast, colon, gastric and prostate carcinomas and pancreatic endocrine tumors), demonstrated a common signature composed of 21 miRNAs differentially expressed in at least three types of tumors. At the top of the list were miR-21, which was expressed excessively in six types of cancer cells, and miR-17-5p and miR-191, which were expressed excessively in five. As the embryological origin of the analyzed tumors was different, the significance of these findings could be that these common miRNAs participate in the fundamental signaling pathways altered in many types of tumors.

In support of the function of these genes in tumorigenesis, it was found that the predicted targets of differentially expressed miRNAs are significantly enriched by those that target suppressors and oncogenes of the known tumor. Also, miR-21, the only miRNA overexpressed in all six types of cancers analyzed, has been shown to directly target the tumor suppressor PTEN, which encodes a phosphatase that inhibits growth and / or survival pathways. PTEN's function is altered in advanced tumors of various types, including breast, ovarian, gastric and prostate.

In some configurations of the subject disclosed herein, a method for diagnosing cancer in an individual is provided. In some configurations, the method comprises providing a biological sample from an individual; isolating cancer-derived exosomes comprising miRNAs from the biological sample;

determining an amount of one or more of the miRNAs; and comparing the amount of one or more miRNAs with one or more levels of miRNA control. The individual can then be diagnosed as having cancer if there is a measurable difference in the amount of one or more miRNAs from the cancer-derived exosomes in the sample compared to one or more levels of control. An unlimited list of exemplary miRNAs is provided that can be measured in Tables 1 and 2. In some configurations, the measured miRNAs are selected from the miRNAs listed in Table 2, and in some particular configurations, the measured miRNAs are selected from the group consisting of miR-21, miR-141, miR-200a, miR-200b, miR -20uc, miR-203, miR-205 and miR-214.

The term cancer refers to all types of cancer or neoplasms or malignant tumors found in animals, including leukemias, carcinomas and sarcomas. Examples of cancers are cancer of the brain, bladder, breast, cervix, colon, head and neck, kidney, lungs, non-small cell lung, melanoma, mesothelioma, ovary, pancreas, prostate, sarcoma, stomach and of the uterus.

r— leucem-ba —- s-ian-if i &amp; a — a — incl-us-ãe — tease — of progressive, malignant diseases of the blood-forming organs, and is generally characterized by uncontrolled proliferation and the development of leukocytes and precursors in the blood and bone marrow. Leukemic diseases include, for example, acute non-lymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, leukemia leukemia, leukocytic leukemia, basophilic leukemia, bloating leukemia, basophilic leukemia , bovine leukemia, chronic myelocytic leukemia, cutis leukemia, embryonic leukemia, eosinophilic leukemia, Gross' leukemia [transmissible murine leukemia], hairy cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, primordial cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphocytic leukemia, lymphocytic leukemia, lymphocytic leukemia, lymphocytic leukemia, lymphocytic leukemia, lymphocytic leukemia , megakaryocytic leukemia, micromieloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, acute myelomonocytic leukemia, plasma cell leukemia, plasmacytic leukemia, promyelocytic leukemia, 10 Rieder cell leukemia, Schilling leukemia, primordial cell leukemia, subleukemic leukemia and subleukemic leukemia.

Carcinoma refers to a malignant new growth composed of epithelial cells that tend to infiltrate the surrounding tissues and trigger metastases. The

T5 carcinomas exemplcrr'e's incinent; —pcrr — exempdrr, -cancoirroma — a-eê-nar-7 acinous carcinoma, adenoid cystic carcinoma, adenocystic carcinoma, adenomatous carcinoma, adrenal cortex carcinoma, alveolar carcinoma, carcinoma of alveolar cells basal cell carcinoma, basaloid carcinoma, basal squamous cell carcinoma, bronchoalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedocarcinoma, carcinoma of the uterus, cribriform carcinoma, carcinoma in armor, skin carcinoma, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, durum carcinoma, embryonic carcinoma, encephaloid carcinoma, encephaloid carcinoma, epithelial, exophytic carcinoma, ex ulcere carcinoma, fibrous carcinoma, gelatiniform carcinoma, gelatinous carcinoma, giant cell carcinoma, gigantocellular carcinoma, glandular carcinoma, granular cell carcinoma, hair matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernefroid carcinoma, infant embryonic carcinoma, in situ carcinoma, intraepidermal carcinoma, intraepidermal carcinoma, intraepidermal carcinoma Krompecher's disease, Kulchitzky cell carcinoma, large cell carcinoma, lenticular carcinoma, lipomatous carcinoma, lymphoepithelial carcinoma, medullary carcinoma, medullary carcinoma, melanotic carcinoma, molle carcinoma, mucinous carcinoma, muciparum carcinoma, mucocellular carcinoma, mucoepidermoid carcinoma, mucosum carcinoma, mucous carcinoma, myxomatous carcinoma, nasopharyngeal carcinoma, carcinoma of the cells,^_you^_ó ~ ± dey ca'r'c'rn'oma papillary, periportal carcinoma, and preinvasive carcinoma, squamous cell carcinoma, pultacea carcinoma, renal kidney cell carcinoma, reserve cell carcinoma, sarcomatode carcinoma, schneiderian carcinoma, cirrhosis carcinoma, scrotum carcinoma, signet ring cell carcinoma, simplex carcinoma, small cell carcinoma, solanoid carcinoma, spheroid cell carcinoma, spindle cell carcinoma, spongiosum carcinoma, squamous carcinoma, squamous cell carcinoma, string carcinoma, telangiectatic carcinoma, telangiectoid carcinoma, transitional cell carcinoma, tuberosum carcinoma, tuberous carcinoma, verrucous carcinoma and yolk sac carcinoma.

The term sarcoma generally refers to a tumor that is composed of a substance similar to embryonic connective tissue, being generally composed of cells closely packed together in a fibrillar or homogeneous substance. Sarcomas include, for example, chondrosarcoma, fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma, Abemethy sarcoma, adipose sarcoma, liposarcoma, soft tissue alveolar sarcoma, ameloblastic sarcoma, chroma sarcoma, sarcoma, botryoid sarcoma, sarcoma Wilms' tumor, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblast sarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic B cell sarcoma, lymphoma, immunoblastic T cell sarcoma, Jensen's sarcoma, Kaposi's sarcoma, sarcoma of

E5-cé-lu ± as-de-KupTíexç-angro'SSU.-rcOmaç-tercosKarcoina, malignant mesenchymal sarcoma, parosteal sarcoma, reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma and telangiectatic sarcoma.

The term melanoma means a tumor that arises from the melanocytic system of the skin and other organs. Melanomas include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, malignant lentigo melanoma, malignant melanoma, nodular melanoma, subungal melanoma and melanoma superficial spread.

In some settings, cancer is a cancer selected from the group consisting of ovarian cancer, cervical cancer, breast cancer, endometrial cancer, colon cancer, prostate cancer, lung cancer, melanoma and pancreatic cancer.

The term biological sample used herein refers to a sample that comprises a biomolecule and / or that is obtained from an individual. Representative biomolecules include, among others, total DNA, RNA, miRNA, mRNA and polypeptides. The biological sample can be used to detect the presence and / or level of expression of a miRNA of interest associated with exosomes derived from cancer. Any cell, group of cells, cell fragment or cell product can be used with the methods of the matter claimed herein, although the biological fluids and organs that would be predicted to contain exosomes derived from cancer and that exhibit differential expression of miRNAs when compared to i5-normal controls ^ —In — a-guma-s — con-f-igurations —; - a — amo-s-tra — biO-ionic — is — a relatively easy-to-obtain biological sample , such as, blood or one of its components. In some configurations, the biological sample comprises milk, blood, serum, plasma, ascites, cystic fluid, pleural fluid, peritoneal fluid, cerebrospinal fluid, tears, urine, saliva, phlegm or combinations thereof.

In some configurations, size exclusion chromatography can be used to isolate cancer-derived exosomes. See, for example, Figures 1 and 2. The size exclusion chromatography techniques are well known. Exemplary and non-limiting techniques are provided in the present Examples. In some configurations, a fraction of empty volume is isolated and comprises the exosomes of interest.

In addition, in some configurations, cancer-derived exosomes can still be isolated after chromatographic separation by centrifugation techniques (of one or more chromatographic fractions), as is generally known in the art. In some 5 configurations, for example, density gradient centrifugation can be used to better isolate exosomes. In some configurations, it may be desirable to further separate isolated exosomes derived from cancer from exosomes from other sources. For example, cancer-derived exosomes can be separated from non-cancer exosomes by immunoabsorbent capture using an anti-cancer antigen antibody. See, for example, Figure 8. Exemplary anti-cancer antigen antibodies include, among others, antibodies to the anti-epithelial cell adhesion molecule (anti-EpCAM), used, for example,

T5 exemptions as indicated in the present Exempitrst

The terms diagnose and diagnosis used herein refer to methods by which those skilled in the art can estimate and even determine whether an individual is suffering from a given disease or problem. Technicians in the subject generally make a diagnosis based on one or more diagnostic indicators, such as, for example, a biomarker (for example, a level of miRNA expression), the quantity (including the presence or absence) of what is indicative of presence, severity or absence of the disease.

Along with the diagnosis, the clinical prognosis of cancer is also an area of ​​great concern and interest. It is important to know the aggressiveness of the cancer cells and the probability of tumor recurrence in order to plan the

1-5 most effective therapy. Some cancers, for example, are treated with several alternative strategies. In certain cases, local-regional and systemic radiation therapy is used, while in other cases surgical intervention and / or chemotherapy are used.

Current treatment decisions for individuals with cancer may be based on (1) the number of lymph nodes involved in the disease, (2) the condition of the cancer marker (s), (3) the size of the primary tumor, and (4) at the stage of the disease in the diagnosis. However, even with these factors, it is not possible to accurately predict the course of the disease for all individuals with cancer. If a more accurate prognosis can be made, the appropriate therapy can be chosen and, in some cases, the least severe therapy for the patient can be chosen. The measurement of exosomal cancer-derived miRNA levels revealed in the present

-for cartregO-rrz-arr o^_s rrrdxvrdutrs in agreement σοτη the fuss of the person who benefits from certain therapies and differentiates themselves from other individuals, in which alternative or additional therapies may be more appropriate. Thus, in some configurations of the matter disclosed herein, a method is provided for categorizing cancer in an individual. In some configurations, the method comprises providing a biological sample from an individual; isolating cancer-derived exosomes comprising micro-RNAs (miRNAs) from the biological sample; determine the quantity of one or more of the miRNAs; and comparing the amount of one or more miRNAs with one or more levels of miRNA control. In these configurations, cancer can be characterized based on a measurable difference in the amount of one or more miRNAs from cancer-derived exosomes compared to one or more levels of miRNA control. In some configurations, the characterization of cancer comprises the determination of a type, class and / or stage of the cancer.

Making a diagnosis or diagnosing, as used herein, is even more consistent than a 5 prognosis, being able to predict a clinical outcome (with or without medical treatment), select an appropriate treatment (or whether the treatment would be effective), or monitor an ongoing treatment and potentially change that treatment, based on the measurement of diagnostic exosomal miRNA diagnostic levels derived from cancer. In addition, in some configurations of the material disclosed herein, multiple determination of the amounts of one or more miRNAs over time can be made to facilitate diagnosis (including prognosis), evaluate the effectiveness of treatment, and / or the progression of a cancer. A temporal change can be made in one or more

-1-5-n-í-veis — d-tag-we-ti-eos- miR-NA — ex-os-somad-s — de-ri-va-do-s — d © —e-â -n-ee-r - (- is-feo — is-amounts of miRNA in the biological sample) to predict a clinical outcome, monitor cancer progression, and / or the effectiveness of administered cancer therapies. In this configuration, for example, it is possible to observe a reduction in the amount of certain miRNAs in the biological sample over the course of a therapy, thus indicating the effectiveness of the treatment.

The material disclosed in the present article still provides in some configurations a method for evaluating the effectiveness and / or progression of cancer treatment in an individual. In some configurations, the method comprises the collection of a series of biological samples over an individual's time period;

isolating cancer-derived exosomes comprising miRNAs from the series of biological samples; determine an amount of one or more of the miRNAs in each of the biological samples in the series; and determining any measurable action in the quantities of one or more miRNAs in each of the biological samples in the series, in order to assess the effectiveness and / or progression of cancer treatment in the individual. Any changes in the amounts of miRNAs measured over time can be used to predict the clinical outcome, determine the initiation or continuation of cancer therapy, and whether ongoing therapy is effectively treating cancer. For example, a first timepoint can be selected before the start of a treatment and a second timepoint some time after the start of the treatment. The miRNA levels can be measured in each sample taken at different points in time, with qualitative and / or quantitative differences being noted. A change in the amounts of one or more of the measured miRNA levels in the

T5 pr'i'rtrerra and the — sagunda — anros-tras — can — co-r-re-laei-onar-with — the prognosis, determining the effectiveness of the treatment, and / or the progression of the disease in the individual.

The correlated and correlating terms as used herein with reference to the use of diagnostic and prognostic miRNA levels associated with cancer, refer to the comparison of the presence or quantity of miRNA levels in an individual with their presence or quantity in individuals who are known to suffer from cancer , or in individuals known to be cancer-free, ie normal individuals or control individuals. For example, a level of one or more miRNAs in the biological sample can be compared to a miRNA level for each of the specific miRNAs tested and determined to correlate with cancer. The miRNA levels of one or more samples are correlated with a diagnosis; that is, those skilled in the art can use miRNA levels (s) to determine whether the individual suffers from cancer and responds accordingly. Alternatively, the sample miRNA levels can be compared to the control miRNA levels known to be associated with a good outcome (for example, the absence of cancer), as a mean level found in a population of individuals normal.

In certain configurations, a diagnostic or prognosis miRNA level is correlated with cancer 10 simply by presence or absence. In other configurations, a threshold level of a diagnostic or prognosis miRNA level can be established, and the level of miRNA in a sample of the individual can be simply compared to the threshold level.

T5 As frrdircardoç — in — al-gumars — eon-figure-çõ-s -&gt; - multiple determinations of one or more diagnostic or prognostic miRNA levels can be made, and a temporal change in levels can be used to determine a diagnosis or prognosis.

For example, specific miRNA levels can be determined at an initial time, and then at a second time. In these configurations, an increase in the miRNA level (s) from an initial time to the second time can be a cancer diagnosis, or a given prognosis. Likewise, a reduction in the miRNA level (s) from an initial time to the second time may be indicative of cancer or a given prognosis. In addition, the degree of change in one or more miRNA levels may be related to the severity of the cancer and / or the progression of the disease and future adverse events.

Those skilled in the art will understand that, while in certain configurations comparative measurements of the same miRNA levels can be made at multiple points in time, it is also possible to measure certain miRNA levels at a 5 timepoint, and second ( s) miRNA levels in a second timepoint, and a comparison of these levels can provide diagnostic information.

The phrase determining the prognosis as used herein refers to methods by which technicians in subject 10 can predict the course or outcome of a disease in an individual.

The term prognosis does not refer to the ability to predict the course or outcome of a disease with 100% accuracy, or even if the given course or outcome is more or less likely to occur based on the presence, absence or levels of a 15 bíOma'rca'dorc — Instead — di-s-so -; —the — fré-en-i-eo-s — ηθ — assun-to — eomp-ree-nd-e-frog-what prognostic term refers to a greater probability of the occurrence of a certain course or result; that is, that a course or result is more likely to occur in an individual who demonstrates a certain condition, when compared 20 with individuals who do not exhibit the condition. For example, in individuals who do not exhibit the condition (for example, who do not express the miRNA level (s) or who express the reduced miRNA level (s)), the probability of a given result ( for example, suffering from cancer) can be very small (for example, &lt;1%) or even nil. In contrast, in individuals who exhibit the condition (for example, who express the miRNA level (s) or who express the miRNA level (s) at a much higher level with respect to the level of control), the likelihood of a given outcome (for example, suffering from a form / stage of cancer) may be high. In certain configurations, a prognosis has about 5% probability of a given expected result, about 7% probability, about 10% probability, about 12%

<td>5 probability,</td><td>fence</td><td>in</td><td> 15%</td><td>in</td><td>probability,</td><td>fence</td><td>in</td><td> 20%</td><td>in</td>

<td>probability,</td><td>fence</td><td>in</td><td> 25%</td><td>in</td><td>probability,</td><td>fence</td><td>in</td><td>or »o.</td><td>in</td>

<td>probability,</td><td>fence</td><td>in</td><td> 40%</td><td>in</td><td>probability,</td><td>fence</td><td>in</td><td> 50%</td><td>in</td>

<td>probability,</td><td>fence</td><td>in</td><td> 60%</td><td>in</td><td>probability,</td><td>fence</td><td>in</td><td> 75%</td><td>in</td>

<td>probability,</td><td>fence</td><td>in</td><td> 90%</td><td>in</td><td>probability or</td><td>fence</td><td>in</td><td> 95%</td><td>in</td>

probability...

Those skilled in the art will understand that the association of a prognostic indicator with a predisposition to an adverse outcome is a statistical analysis. For example, miRNA levels (for example, quantity of one or more miRNAs in a sample) greater or less than a level of co'n't'roi “eç in^—ai-guma-s · configurations, may signal that an individual is more likely to suffer from cancer than individuals with a level below or equal to the level of control, as determined by a level of statistical significance. In addition, a change in the miRNA level (s) from baseline levels may reflect an individual's prognosis, and the degree of change at the marker level may be related to the severity of adverse events. Statistical significance is generally determined by comparing two or more populations, and by determining a confidence interval and / or a p-value. See, for example, Dowdy and Wearden, Statistics for Research, John Wiley &amp; Sons, New York, 1983, incorporated into presence by reference in its entirety. The exemplary confidence intervals for this subject are 90%, 95%, 97.5%, 98%, 99%,

99.5%, 99.9% and 99.99%, while the exemplary p-values ​​are 0.1,

0.05, 0.025, 0.02, 0.01, 0.005, 0.001 and 0.0001.

In other configurations, a limit level of level change of one or more prognostic or diagnostic miRNA levels can be established, and the degree of change in the level of the indicator in a biological sample can be simply compared to the limit degree of level change. A preferred limit change in the miRNA level (s) of the material disclosed herein is about 5%, about 10%, about 15%, about 20%, about

25%, about 30%, about 50%, about 60%, about 75%, about 100%, and about 150%. In still other configurations, a nomogram can be established, whereby the level of a prognostic or diagnostic indicator can be directly related to an associated disposition directed to a given result. Technicians in the subject are familiar with the use of such nomograms that relate two numerical values ​​with the understanding that the uncertainty in this measurement is the same as the uncertainty in the concentration of the marker, because the measurements of the individual sample are referenced, not being averages of population.

The identity and relative quantity of miRNAs in the sample can be used to provide miRNA profiles for a given sample. A miRNA profile for a sample includes information about the identities of the miRNAs contained in the sample, the quantitative levels of miRNAs contained in the sample, and / or changes in the quantitative levels of miRNAs relative to another sample. For example, a sample miRNA profile includes information about the identities, quantitative levels and / or changes in the quantitative levels of miRNAs associated with a given cancer.

Still with regard to the diagnostic methods of the matter disclosed herein, a preferred individual is a vertebrate individual. A preferred vertebrate is warm-blooded;

a preferred warm-blooded vertebrate is a mammal. A preferred mammal is preferably human. As used herein, the term individual includes both human and animal individuals. Thus, veterinary therapeutic uses are provided according to the material disclosed herein.

Thus, the matter disclosed in the present provides the diagnosis of mammals as humans, as well as mammals of importance because they are in danger of extinction, such as Siberian tigers; of economic importance, such as animals raised on farms for human consumption; and / or animals of social importance to humans, such as pets or that are in zoos. Examples of these animals include, but are not limited to:

carnivores like cats and dogs; pigs, including pigs, capons and wild boars; ruminants and / or ungulates such as cattle, oxen, sheep, giraffes, deer, goats, bison and camels; and horses. The treatment of birds is also provided, including the treatment of types of birds in danger of extinction and / or kept in zoos, as well as edible birds, and more particularly domesticated birds, that is, domestic birds, such as turkeys, chickens, ducks, geese, guinea fowl and the like, as they are also of economic importance to humans. Thus, breeding treatment is also provided, including, among others, domesticated pigs, ruminants, ungulates, horses (including racehorses), poultry and the like.

As indicated above, the material disclosed herein provides for the determination of the amount of exosomal cancer-derived miRNAs correlated with cancer within an individual's biological fluids and, in particular, an individual's serological samples, such as blood. This gives an advantage to testing biological samples that are easily obtained from an individual. The amount of one or more miRNAs of interest in the biological sample can then be determined using any number of methodologies generally known in the art and compared to the levels of miRNA control.

The quantity of one or more miRNAs determined refers to a qualitative (for example, present or not in the measured sample) and / or quantitative (for example, how much is present) measurement of one or more miRNAs. The level of control is an amount (including qualitative presence or absence) or range of quantities of one or more miRNAs found in a comparable biological sample in individuals who do not suffer from cancer. As a non-limiting example of calculating the level of control, the amount of one or more miRNAs of interest present in a normal biological sample (for example, blood) can be calculated and extrapolated to all individuals.

An exemplary methodology for measuring the miRNA levels of exosomes in a biological sample is the microarray technique, which is a powerful tool applied to studies of gene expression. The technique provides many polynucleotides with sequential information known as probes to find and hybridize to the complementary strands in a sample, so as to capture the complementary strands by selective binding. Figures and 8 flow diagrams of exemplary protocols for the isolation and measurement of miRNAs with exosomal derivation by microarray.

The term selective binding as used herein refers to a measure of a probe's ability to hybridize to a target polynucleotide with specificity. Thus, the probe comprises a complementary, or essentially complementary, polynucleotide sequence in at least part of the target polynucleotide sequence. The nucleic acid sequences that are complementary are those with base pairing according to standard Watson-Crick complementarity rules. As used herein, the term complementary sequences means sequences of nucleic acids that are substantially complementary, as can be assessed by the same nucleotide comparison indicated above, or as defined as being able to hybridize the nucleic acid segment in question under relatively strict conditions , as described above. A particular example of a contemplated complementary nucleic acid segment is an antisense oligonucleotide. With respect to the probes disclosed herein having binding affinity with miRNAs, the probe can be 100% complementary to the target polynucleotide sequence. However, the probe does not necessarily have to be completely complementary to the target polynucleotide along the entire length of the target polynucleotide as long as the probe can specifically bind to the target polynucleotide and capture it from the sample.

The hybridization of the nucleic acid will be affected by conditions such as saline concentration, temperature or organic solvents, in addition to the base composition, length of the complementary strips and the number of non-combinations of nucleotide base between the hybridizing nucleic acids, as will be readily appreciated by those skilled in the art. . Rigid temperature conditions generally include temperatures greater than 30 ° C, typically greater than 37 ° C and preferably greater than 45 ° C. Rigid saline conditions will normally be less than 1,000 mM, typically less than 500 mM and preferably less than

200 mM. However, the combination of parameters is much more important than the measurement of any single parameter. The determination of suitable hybridization conditions to identify and / or isolate sequences containing high levels of homology is well known in the art. For the purpose of specifying high stringency conditions, the preferred conditions are a salt concentration of about 200 mM and a temperature of about 45 ° C.

The data research work is completed by bioinformatics, including scanning chips, signal acquisition, image processing, normalization, statistical treatment and data comparison, as well as path analysis. Thus, the microarray technique can profile hundreds and thousands of polynucleotides simultaneously with high performance. The microarray profiling analysis of mRNA expression has successfully provided valuable data for gene expression studies in basic research. And the technique has also been put into practice in the pharmaceutical industry and in clinical diagnosis. With increasing amounts of miRNA data becoming available, and with the accumulation of evidence of the importance of miRNA in genetic regulation, the microarray technique becomes useful in high-performance miRNA studies.

The analysis of miRNA correlated with cancer can be done separately or simultaneously with multiple polynucleotide probes within a test sample. For example, multiple probes can be combined in one test for efficient processing of multiple samples and to provide potentially greater diagnostic and / or prognostic accuracy. In addition, those skilled in the art would recognize the value of multiple sample tests (for example, at successive points in time) from the same individual. These serial sample tests can allow identification of changes in miRNA levels over time. Increases or decreases in miRNA levels, as well as no change in levels, can provide useful information about the condition of the disease.

In some configurations, a panel can be constructed that consists of polynucleotide probes that selectively bind exosomal cancer-derived miRNAs correlated to one or more cancers to provide relevant information related to the diagnosis or prognosis of cancer and the treatment of individuals with cancer. This panel can be built, for example, using 1, 2, 3, 4, 5, 6, 7,

8, 9, 10, 15, 20, 30, 40, 50, 75, 100, 150, 200, 250, 300, 400,

500 or 1,000 individual polynucleotide probes. Analysis of a single probe or subset of probes comprising a larger probe panel can be done by those skilled in the art in order to optimize sensitivity or clinical specificity in various clinical settings. These include, among others, outpatient units, urgent treatments, critical treatments, intensive care, monitoring units, individual hospitalization, individual outpatient, doctor's office, medical clinic and health screening units. In addition, those skilled in the art can use a single probe or a subset of other probes, comprising a larger probe panel in combination with an adjustment of the diagnostic limit in each of the aforementioned units to improve clinical sensitivity and specificity. The clinical sensitivity of a trial is defined as the percentage of those with the disease correctly predicted by the trial, and the specificity of a trial is defined as the percentage of those without the disease correctly predicted by the trial.

In some configurations, determining the amount of one or more miRNAs comprises labeling one or more miRNAs. The labeled miRNAs can then be captured with one or more polynucleotide probes that selectively bind one or more miRNAs.

As used herein, the terms label and marked refer to the attachment of a medium capable of detection by spectroscopic, radiological or other methods to a molecular probe. Thus, the terms label or marked refer to the incorporation or attachment, optionally covalently or non-covalently, of a detectable marker within / over a molecule, such as a polynucleotide. Various methods of labeling polypeptides that can be used are known in the art. Examples of polynucleotide labels include, but are not limited to, the following: radioisotopes, fluorescent labels, heavy atoms, enzyme labels or reporter genes, chemiluminescent groups, biotinyl groups, predetermined polypeptide epitopes recognized by a secondary reporter (e.g. zipper pair sequences from leucine, antibody binding sites, metal binding domains, epitope identifiers, etc.). In some configurations, the labels are attached by spacer arms of various lengths to reduce potential steric hindrance.

Analysis of miRNA levels can also be performed using polynucleotide probes in various physical formats. For example, microtiter or automation plates can be used to facilitate the processing of a large number of test samples. Alternatively, simple samples could be developed to facilitate immediate treatment and diagnosis within a time frame.

In some configurations, the plurality of polynucleotide probes are attached to a substrate. In some configurations, the substrate comprises a plurality of addresses. Each address can be associated with at least one of the polynucleotide probes in the array. An array is addressable when it has multiple regions of different media (e.g. different polynucleotide sequences) so that a region (ie, a feature or spot in the array) at a particular location (ie, an address) in the array will detect a particular target or class of targets (although a feature may incidentally detect not targets of that characteristic). Arrangement characteristics are commonly, but need not be, separated by intervention spaces. In the case of an array, the miRNA target can be referred to as a medium in a mobile (commonly fluid) phase, to be detected by probes (target probes) that are attached to the substrate in several regions.

Arrangements of biopolymers (eg polynucleotide microarrays) can be manufactured by depositing previously obtained biopolymers (such as from synthesis or from natural sources) on a substrate, or by in situ synthesis methods.

Methods for depositing the obtained biopolymers include, among others, loading and then touching a pin or capillary on a surface, as described in U.S. Patent No. 5,807,522 or jet deposition. pulse, as for an inkjet head, as described in PCT publications WO 95/25116 and WO 98/41531 and elsewhere.

In situ manufacturing methods include those described in

U.S. Patent No. 5,449,754 for the synthesis of peptide arrays and in U.S. Patent No. 6,180,351 and

WO 98/41531 and the references cited therein for polynucleotides, and may also use pulse jets to deposit reagents. Further details of the manufacture of biopolymer arrangements by deposit, whether biopolymers previously obtained or by the in situ method are disclosed in the United States Patents

We. 6,242,266, 6,232,072, 6,180,351 and 6,171,797. In the manufacture by depositing previously obtained biopolymers or by in situ methods, typically each region on the surface of the substrate on which the array will be or was formed (array regions) is completely exposed to one or more reagents. For example, in either method, the arrangement regions will generally be exposed to one or more reagents for the formation of a suitable layer on the surface that binds to both the substrate and the biopolymer or to the biomonomer. In on-site manufacturing, the arrangement regions will also normally be exposed to oxidation, deblocking and optional capping reagents. Similarly, particularly in the manufacture by depositing previously obtained biopolymers, it may also be desirable to expose the arrangement regions to a suitable blocking reagent for blocking sites on the surface where there are no non-specific binding characteristics to the target.

The determination of the amount of exosomal miRNAs derived from cancer may alternatively, or in addition to microarray analysis, comprise the use of real-time polymerase chain reaction (PCR), for example, in the manner disclosed in detail in the present Examples. Real-time PCR (RT-PCR) can provide fast and accurate data such as the presence and quantity of miRNAs in a sample. Figure 2 provides a flowchart of an exemplary protocol for the isolation and measurement of exosomal miRNAs by RT-PCR. Further details of the exemplary methodologies are given in the present Examples.

In some configurations of the material disclosed herein, a method is provided for diagnosing potential adverse pregnancy outcomes in an individual. The methodology mentioned in detail for the isolation of exosomes comprising miRNAs and the determination of the amount of miRNAs can also be applied for the quantification of certain miRNAs associated with adverse pregnancy outcomes, with some modifications that will now be described.

In order to predict adverse pregnancy outcomes, circulating exosomes obtained from the placenta can be isolated from the biological sample, for example, blood or its components. The placenta, although derived from the fetus, is the only fetal tissue actually in contact with the maternal system. Thus, the exosomes produced by the placental cells can circulate in the mother's blood flow. For the isolation of exosomes derived from the placenta, both anti-EpCAM antibodies (as used for the isolation of tumor exosomes) and type i alkaline phosphatase antibodies can be used

antiplacental (PLAP) attached to magnetic beads (see, eg

Figure 8).

For example, in some configurations, the method comprises providing a biological sample from an individual and isolating exosomes comprising micro-RNAs (miRNAs) from the biological sample. An amount of one or more of the miRNAs is then determined and compared with one or more levels of miRNA control. The individual can then be diagnosed as being at risk of an adverse pregnancy outcome if there is a measurable difference in the amount of one or more miRNAs from the exosomes when compared to one or more levels of miRNA control. In some settings, the result of adverse pregnancy is a disorder selected from the group consisting of preeclampsia, premature birth (eg. delivery before 32 weeks of gestation), premature rupture of membranes, restriction of intrauterine growth and recurrent pregnancy loss.

The practice of the subject disclosed herein may employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA and immunology, which are inserted in the technique. These techniques are fully explained in the literature. See, for example,

Molecular Cloning A Laboratory Manual (1989), 2nd Ed., Ed. by

Sambrook, Fritsch and Maniatis, eds., Cold Spring Harbor

Laboratory Press, Chapters 16 and 17; US Patent No.

4,683,195; DNA Cloning, Volumes I and II, Glovide, ed., 1985;

Oligonucleotide Synthesis, MJ Gait, ed., 1984; Nucleic Acid

Hybridization, D. Hames &amp; SJ Higgins, eds., 1984; Transcription and Translation, BD Hames &amp; SJ Higgins, eds., 1984; Culture

Of Animal Cells, RI Freshney, Alan R. Liss, Inc., 1987;

Immobilized Cells And Enzymes, IRL Press, 1986; Perbal (1984), A

Practical Guide To Molecular Cloning; See Methods In Enzymology (Academic Press, Inc., NY); Gene Transfer Vectors For Mammalian

Cells, JH Miller and Μ. P. Calos, eds., Cold Spring Harbor

Laboratory, 1987; Methods In Enzymology, Vols. 154 and 155, Wu et al., Eds., Academic Press Inc., NY; ImmunoChemical Methods In

Cell And Molecular Biology (Mayer and Walker, eds., Academic

Press, London, 1987; Handbook Of Experimental Immunology, Volumes

I-IV, DM Weir and CC Blackwell, eds., 1986.

EXAMPLES

The following Examples have been included to illustrate the modes of matter disclosed herein. In the light of the present disclosure and at the general level of the current technique, those skilled in the art will appreciate that the following Examples have only the exemplary function and that numerous changes, modifications and alterations can be made without departing from the scope of the matter disclosed herein.

The material revealed in the present reveals that miRNA can be found and isolated from exosomes in biological fluids. Isolated miRNA can be used as a diagnostic tool for cancer and adverse pregnancy outcomes. The present Examples support these applications.

MATERIALS AND METHODS FOR EXAMPLES 1-5

Patient samples and cell lines

These Examples used as exemplary biological fluids derived from serum of women diagnosed with serous papillary adenocarcinoma of the ovary (n = 50; n = 10 for stage

I, n = 10 for stage II, n = 20 for stage III and n = 10 for stage

IV), women of the same age with benign ovarian adenoma (n = 10), and women of the same age without evidence of ovarian disease (n = 10). Controls were selected, patients with benign ovarian disease and ovarian cancer in stages III and IV based on the age combination of patients with early stage ovarian cancer. These Examples also include data from investigations of primary tumor cell cultures, established from 6 women with Stage IIIc ovarian cyst adenocarcinoma and their corresponding pre-surgical serum samples. All of these materials were obtained with informed consent approved by University Human Studies

University of Louisville Committee.

Cell cultures of primary ovarian tumors were made in our laboratory and named UL-1, UL-2, UL-3, UL-6, UL-B and UL-O. UL-2 and UL-3 were obtained from hereditary ovarian cancers, while UL-1, UL-6, UL-B and

UL-0 were obtained from spontaneous cancers. These ovarian tumor cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (by ultrafiltration) free of exosomes, 0.1 mM non-essential amino acids, 1 mM sodium pyruvate, 200 mM L-glutamine, 100 mg / ml streptomycin and 100 IU / ml penicillin in CO atmosphere₂ humidified to 5%. Cell viability was assessed by excluding trypan blue and all cultures used were&gt; 95% viable.

Isolation of circulating exosomes

Tumor-derived exosomes were specifically isolated by a modified magnetic activation cell choice procedure (MACS), using antiepithelial cell adhesion molecule (EpCAM). Our previous studies have shown that exosomes from epithelial tumors express

EpCAM on their surfaces and can be used for their selective insulation. Serum samples (2.5 ml) from normal controls, patients with benign diseases and patients with early stage ovarian cancer were incubated with anti-EpCAM coupled to magnetic microcounts (50 μΐ). They were mixed and incubated for 2 hours at 4 ° C. An LD microcolumn was placed in the magnetic field of a MACS Separator and the column was rinsed with

500 μΐ of Tris-saline buffer (TBS). The immune magnetic complexes were applied to the column and the unbound (unmarked) material passed through and was discarded. The column was washed four times with 500 μΐ of TBS. The specifically selected exosomes were recovered by removing the column from the separator and placing them in a collection tube. TBS (1 ml) was added to the column, obtaining the magnetically marked exosomes by applying the plunger provided with the column. The isolated exosomes / micro accounts were diluted in IgG elution buffer (Pierce Chemical Co, Rockford, IL) and the complex was centrifuged at 10,000 rpm to separate the micro accounts from the exosomes (supernatants). The supernatant was then centrifuged at 10000g for 1 hour at 4 ° C. The pelleted exosomes were resuspended in 250μ1 in phosphate buffered saline (PBS) and tumor-derived exosomes were tested for total protein. The amount of protein was determined using the Bradford microassay method (Bio-Rad Laboratories, Hercules,

CA), using bovine serum albumin (BSA) as standard.

Transmission electronic microscopy

For transmission electron microscopy, the pelleted exosomes were fixed in 2.5% (w / v) of glutaraldehyde in PBS, dehydrated and integrated in Epon. Ultra-thin sections (65 nm) were cut and stained with uranyl acetate and Reynold's lead citrate. The sections were examined using a Jeol 1210 transmission electron microscope.

MiRNA isolation and profiling

Total RNA was isolated from tumor cells and exosomes using the mirVana miRNA isolation kit according to the manufacturer's instructions (Ambion, Austin, TX). The quality of the RNA, the production and the size of the miRNA fractions were analyzed using the Agilent 2100 Bioanalyzer (Agilent

Technologies, Foster City, CA). Isolated miRNAs were labeled with Cy3 at the 3 'end using the mirVana miRNA Array Kit

Labeling Kit (Ambion) and Post Labeling Reactive Dye kit (Amersham

Bioscience, Pittsburgh, PA). The microRNA profiling was done in duplicate by Ocean Ridge Biosciences (Jupiter, FL) using microarrays containing probes for 467 mature human miRNAs. This analysis used miRNA arrays specially developed covering the 467 miRNAs present at the Sanger Institute mirBASE v9.0, which consists of 35-44-mer oligonucleotides, manufactured by

Invitrogen and marked in duplicate. After hybridization, miRNA arrays were scanned using a GenePix 4000A array scanner (Axon Instruments, Union City, CA) and the raw data normalized and analyzed using GeneSpring 7.0

Software (Silicon Genetics, Redwood City, CA). Normalization was done by expressing each miRNA replicate in relation to the control miRNA (Ambion) added to each sample, allowing comparison between arrangements. The limit and 95 percentile of negative controls (TPT95) were calculated based on the hybridization signal from the negative control probes including: 38 decomposed and mixed control probes and 87 non-conserved C. elegans probes. To define the sensitivity, NCode synthetic miRNA with a mass ratio of 1 / 500,000 was added to the labeling reactions, and the signal strength was detected. For specificity, perfect combination probes for miR-93, miR-27a and miR-152 and 2 were not combined. The 2 unpaired base pair probes showed a signal below or in TPT95 in all arrays.

To assess the stability of the exosomal profiling with storage and manipulation, serum was collected from patients with ovarian cancer and divided into four 4 ml samples. The tumor exosomes were immediately isolated from the first aliquot by MACS procedure and the total RNA was isolated and stored at -70 ° C until all samples were isolated. The remaining serum samples were stored at 4 ° C for subsequent isolation of exosomes. The tumor exosomes were isolated from the second aliquot after 24 hours, from the third aliquot after 48 hours and from the fourth aliquot after 96 hours at 4 ° C. The RNA was isolated from each exosome preparation and stored. In a similar study, 3 other serum aliquots were stored at -70 ° C for 28 days, prior to isolation of exosomes and RNA to mimic the use of specimens in a bank.

General Statistical Considerations

The data were analyzed using the statistical software package, SAS9.1 (SAS Institute, Cary, NC). The levels of circulating exosomes for each group of patients were defined as mean ± standard deviations of at least two separate experiments done in triplicate. The comparisons between these groups were made by one-way ANOVA, followed by Tukey's post-multiple comparisons comparing each population.

The relative quantification of miRNA expression was calculated by the 2-AACt method (Applied Biosystems User Bulletin No. 2), the data being analyzed as log10 of the relative quantity (RQ) of the target miRNA, normalized with respect to the control miRNA added to each sample, allowing comparisons between arrangements. The miRNA distributions and correlations were calculated together with the confidence intervals of each subarray. Statistical significance was established at p &lt;0.05.

EXAMPLE 1

PRESENCE OF POSITIVE EPCAM CIRCULATING EXOSOMES

IN WOMEN WITH BENIGNA AND MALIGNA OVARIAN DISEASE

EpCAM-positive exosomes were isolated specifically using magnetic anti-EpCAM beads, and these circulating exosomes were tested for total protein and plotted for disease stage (Figure 3A). The levels of EpCAM-positive exosomes in normal volunteers of the same age (control) were 0.039 ± 0.030mg / ml of exosomal protein, which represented the background of the assay. Patients diagnosed with benign ovarian disease had 0.149 ± 0.065mg / ml of exosomal protein, which was significantly elevated compared to controls. All patients diagnosed with ovarian cancer exhibited significantly high levels of EpCAMpositive exosomes (compared to benign disease or controls).

Women with Stage I ovarian cancer exhibited

0.320 ± 0.056mg / ml of circulating exosomal protein, which was significantly higher for both controls and benign disease (p &lt;0.01). Circulating exosome levels increased with stage progression, with Stage II cancer having

0.640 ± 0.053mg / ml, Stage III having 0.995 ± 0.084mg / ml and Stage IV showing 1.42 ± 0.228mg / ml. The levels of exosomes associated with these three stages were significantly higher than in women with benign disease or controls (p &lt;0.001).

The resulting fractions were also analyzed by electron microscopy, which demonstrated vesicular structures characteristic of exosomes (Figure 3B). The exosomal nature of this material was further confirmed by the presence of tetraspanins, class I antigens, alkaline phosphatase of the placental type by Western Blot.

EXAMPLE 2

ASSOCIATION OF SMALL RNAs WITH EXOSOMES

TUMOR DERIVATIVES

To identify whether these isolated exosomes contain small RNAs, they are examined using a Bio-Analyzer 2100 (Figure 4). This analysis identified the presence of a significant population of small RNAs in the absence of 18S RNA and

28S, generally seen in cell-derived RNA. This material was then used for miRNA profiling.

EXAMPLE 3

PROFILE OF EXOSOME DERIVED MIRN

CELL DERIVED VERSUS

The presence and levels of cell-specific miRNAs as derived from exosomes were determined using microarray analysis (Figure 1) with probes for 467 miRNAs. Exemplary results are shown in Table 1. The miRNA profiles of our ovarian tumors confirmed the previously reported changes (lorio et al.,

2007). In addition, we demonstrate that among the 467 miRNAs, 218 were above the normalized limit, calculated based on the 95² percentile of the negative control probe signal in both cells and exosomes (Table 2). Of the 218 positive miRNAs, the levels of 175 were not significantly different between ovarian tumor cells and their corresponding exosomes. By comparison, they were present in a greater proportion in the cells, while 31 were present in high levels within the exosomes.

Previously, specific miRNAs have been shown to be overexpressed in human ovarian cancer cells (miR-21, miR-141, miR-200a, miR-200c, miR-200b, miR203, miR-205 and miR-214). To correlate these findings with the material derived from the exosomes, fractions of RNA were isolated from the cells of the original tumor and from the circulating tumor exosomes of the same patients (Figure 5). Using microarray analysis, comparisons between tumor-derived miRNA profiles and exosomal miRNAs derived from peripheral blood indicated that they were not significantly different. In addition, the levels of the 5 tumor-derived miRNA profiles demonstrated a strong correlation with the levels of exosomal miRNAs derived from peripheral blood (for miR-21, r = 0.77; miR-141, r = 0.88; miR- 200a, r = 0.76; miR-200b, r = 0.85; miR-200c, r = 0.83; miR-203, r = 0.85; miR-205, r = 0.91; and miR214 , r = 0.71).

TABLE 1

QUANTITATIVE COMPARISON OF MYRNA IN EXOSOMES

CIRCULANTS AND TUMOR CELLS IN INDIVIDUALS WITH CANCER *

<td></td><td></td><td>Patl Ex</td><td>Cells Patl</td><td>Pat2 Ex</td><td>Cells Pat2</td>

<td>Name</td><td>ID</td><td>866A</td><td>866B</td><td>866C</td><td>866D</td>

<td>hsa-miR-296</td><td> 1098</td><td> 5, 05</td><td> 4,33</td><td> 4,24</td><td> 4,79</td>

<td>hsa-miR-330</td><td> 1002</td><td> 2,98</td><td> 3,09</td><td> 4,1</td><td> 1,08</td>

<td>hsa-miR-20a</td><td> 1007</td><td> 11,46</td><td> 11,35</td><td> 12</td><td> 11,93</td>

<td>hsa-miR-28</td><td> 1024</td><td> 9,4</td><td> 10,05</td><td> 9, 19</td><td> 9,23</td>

<td>hsa-miR-302c</td><td> 1032</td><td> -0,58</td><td> 3,08</td><td> 3, 5</td><td> 1,08</td>

<td>hsa-miR-302a</td><td> 1036</td><td> 2,17</td><td> 3, 66</td><td> 3,47</td><td> 4,33</td>

<td>hsa-miR-214</td><td> 1057</td><td> 6, 58</td><td> 3, 93</td><td> 6, 17</td><td> 2,99</td>

<td>hsa-miR-99b</td><td> 1063</td><td> 9, 59</td><td> 10,08</td><td> 9,86</td><td> 9, 16</td>

<td>hsa-miR-99a</td><td> 1068</td><td> 3,81</td><td> 4,53</td><td> 7,34</td><td> 6,46</td>

<td>hsa-miR-lOa</td><td> 1072</td><td> 10,1</td><td> 10,76</td><td> 9, 63</td><td> 9,55</td>

<td>hsa-let-7d</td><td> 1085</td><td> 12,53</td><td> 13, 32</td><td> 12,65</td><td> 12,5</td>

<td>hsa-miR-138</td><td> 1089</td><td> 5,37</td><td> 5,26</td><td> 4,18</td><td> 3, 61</td>

<td>hsa-miR-140</td><td> 1106</td><td> 3,23</td><td> 4,3</td><td> 2,01</td><td> 0,58</td>

<td>hsa-miR-23a</td><td> 1114</td><td> 14,51</td><td> 15, 08</td><td> 14,99</td><td> 14,78</td>

<td>hsa-miR-215</td><td> 1122</td><td> 0,71</td><td> 1,79</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-183</td><td> 1127</td><td> 9, 08</td><td> 9, 63</td><td> 8,99</td><td> 8,9</td>

<td>hsa-miR-32</td><td> 1135</td><td> 2</td><td> 2, 49</td><td> 2,42</td><td> 0,58</td>

<td>hsa-miR-25</td><td> 1139</td><td> 11,34</td><td> 11,3</td><td> 12,23</td><td> 12,01</td>

<td>hsa-miR-218</td><td> 1143</td><td> 2,71</td><td> 3,37</td><td> 4,61</td><td> 5, 33</td>

<td>hsa-miR-107</td><td> 1163</td><td> 11,68</td><td> 12,18</td><td> 11,29</td><td> 11,31</td>

<td>hsa-miR-145</td><td> 1168</td><td> 1,74</td><td> 2,38</td><td> 3,47</td><td> 1,38</td>

<td>hsa-miR-181a</td><td> 1172</td><td> 11,9</td><td> 12,62</td><td> 11,35</td><td> 11,15</td>

<td>hsa-miR-125a</td><td> 1193</td><td> 12,34</td><td> 13, 07</td><td> 11,67</td><td> 11,84</td>

<td>hsa-miR-222</td><td> 1198</td><td> 12,37</td><td> 12,53</td><td> 11,77</td><td> 10,99</td>

<td>hsa-miR-372</td><td> 1105</td><td> -0,58</td><td> 3,08</td><td> 2,51</td><td> 1,08</td>

<td>hsa-miR-142-3p</td><td> 1253</td><td> 2,72</td><td> 3,06</td><td> 4,59</td><td> 3, 91</td>

<td>hsa-miR-198</td><td> 1258</td><td> 4,2</td><td> 3, 92</td><td> 3,32</td><td> 3, 67</td>

<td>hsa-miR-196a</td><td> 1263</td><td> 4,78</td><td> 5,07</td><td> 3,42</td><td> 4,04</td>

<td>hsa-miR-16</td><td> 1272</td><td> 12,28</td><td> 12,05</td><td> 12, 98</td><td> 12, 6</td>

<td>hsa-miR-200a</td><td> 1287</td><td> 8,29</td><td> 8,72</td><td> 7,17</td><td> 7,44</td>

<td>hsa-miR-18a</td><td> 1292</td><td> 6,41</td><td> 6, 66</td><td> 7,98</td><td> 8,5</td>

<td>hsa-miR-101</td><td> 1297</td><td> 4,62</td><td> 4,87</td><td> 5,55</td><td> 6, 01</td>

<td>hsa-miR-195</td><td> 1311</td><td> 6, 09</td><td> 6, 58</td><td> 6,03</td><td> 6, 43</td>

<td>hsa-miR-136</td><td> 1203</td><td> 3, 69</td><td> 3,77</td><td> 3,52</td><td> 3, 65</td>

<td>hsa-miR-153</td><td> 1225</td><td> 1,71</td><td> 2,08</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-31</td><td> 1227</td><td> 8,97</td><td> 9, 49</td><td> 9,6</td><td> 9,32</td>

<td>ii 3 &lt;3. ~ ITil R ”3 3</td><td>1 Ο Ί Λ J_ SC 1 “1</td><td> 2,01</td><td> 3, 66</td><td>Ί C, Q o / or</td><td>Q Π R Ό ft</td>

<td>hsa-miR-147</td><td> 1278</td><td> 4,65</td><td> 2,95</td><td> 3,47</td><td> 3, 17</td>

<td>hsa-miR-106b</td><td> 1282</td><td> 9, 47</td><td> 9, 19</td><td> 10, 59</td><td> 10,38</td>

<td>hsa-miR-212</td><td> 1288</td><td> 2,67</td><td> 1,81</td><td> 2,89</td><td> 3,75</td>

<td>hsa-miR-15a</td><td> 1312</td><td> 9, 92</td><td> 10, 12</td><td> 10,79</td><td> 11,06</td>

<td>hsa-miR-135b</td><td> 1331</td><td> 4,51</td><td> 4,03</td><td> 4,42</td><td> 4,07</td>

<td>hsa-miR-92</td><td> 1335</td><td> 12,29</td><td> 12,63</td><td> 12, 62</td><td> 12,26</td>

<td>hsa-miR-342</td><td> 1349</td><td> 9, 14</td><td> 9, 62</td><td> 8,36</td><td> 8,39</td>

<td>hsa-miR-205</td><td> 1368</td><td> 6, 15</td><td> 5,74</td><td> 15,25</td><td> 14,33</td>

<td>hsa-miR-150</td><td> 1385</td><td> 4,02</td><td> 2,87</td><td> 3,33</td><td> 1,38</td>

<td>hsa-miR-149</td><td> 1392</td><td> 6,23</td><td> 5, 65</td><td> 5, 92</td><td> 4,79</td>

<td>hsa-let-7b</td><td> 1431</td><td> 12,73</td><td> 12,43</td><td> 13, 73</td><td> 14,06</td>

<td>hsa-miR-370</td><td> 1445</td><td> 4,14</td><td> 3,7</td><td> 4,37</td><td> 2,58</td>

<td>hsa-miR-20 6</td><td> 1449</td><td> 5,22</td><td> 4,34</td><td> 5,58</td><td> 4,91</td>

<td>hsa-miR-128a</td><td> 1350</td><td> 7,54</td><td> 7,9</td><td> 8,06</td><td> 8,01</td>

<td>hsa-miR-197</td><td> 1487</td><td> 11,05</td><td> 9, 76</td><td> 10, 03</td><td> 9,28</td>

<td>hsa-miR-182</td><td> 1506</td><td> 10,11</td><td> 10,89</td><td> 10, 09</td><td> 10,41</td>

<td>hsa-miR-553</td><td> 1750</td><td> 2,77</td><td> 2,95</td><td> 4,05</td><td> 3, 17</td>

<td>hsa-miR-519d</td><td> 1766</td><td> 2,17</td><td> 3,28</td><td> 0,51</td><td> 3,04</td>

<td>hsa-miR-520g</td><td> 1770</td><td> -0,58</td><td> 1,9</td><td> 3,51</td><td> 2,49</td>

<td>hsa-miR-520e</td><td> 1774</td><td> -0,58</td><td> 1,79</td><td> 0,51</td><td> 3,75</td>

<td>hsa-miR-329</td><td> 1778</td><td> 2,67</td><td> 2,08</td><td> 3,32</td><td> 2,91</td>

<td>hsa-miR-634</td><td> 1782</td><td> 6, 69</td><td> 2,64</td><td> 3,32</td><td> 1,08</td>

<td>hsa-miR-614</td><td> 1786</td><td> 1,21</td><td> -0,01</td><td> 1,31</td><td> 2,91</td>

<td>hsa-miR-223</td><td> 1467</td><td> 2,71</td><td> 2,49</td><td> 4,17</td><td> 3,17</td>

<td>hsa-miR-485-5p</td><td> 1863</td><td> 4,27</td><td> 2,49</td><td> 2,6</td><td> 0, 58</td>

<td>hsa-miR-369-5p</td><td> 1867</td><td> 2</td><td> 1,49</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-575</td><td> 1871</td><td> 2,75</td><td> 2,69</td><td> 4,36</td><td> 3,75</td>

<td>hsa-miR-663</td><td> 1891</td><td> 5, 41</td><td> 5</td><td> 6,17</td><td> 5,76</td>

<td>hsa-miR-520f hsamiR-520c</td><td> 1802</td><td> 1,61</td><td> 1,79</td><td> 2,97</td><td> 3, 15</td>

<td>hsa-miR-382</td><td> 1806</td><td> 4,48</td><td> 4,14</td><td> 4,04</td><td> 3,25</td>

<td>hsa-miR-655</td><td> 1920</td><td> 1,21</td><td> 2,3</td><td> 2,1</td><td> 3,17</td>

<td>hsa-miR-545</td><td> 1932</td><td> 2,5</td><td> 2,66</td><td> 2,92</td><td> 3,58</td>

<td>hsa-miR-502</td><td> 1940</td><td> 3,46</td><td> 4,16</td><td> 4,99</td><td> 3,75</td>

<td>hsa-miR-200a *</td><td> 1952</td><td> 5,35</td><td> 5,86</td><td> 3,42</td><td> 2,25</td>

<td>hsa-miR-640</td><td> 1956</td><td> 2,24</td><td> -0,51</td><td> 2,51</td><td> 0, 58</td>

<td>hsa-miR-514</td><td> 1972</td><td> 2</td><td> 2,95</td><td> 1,01</td><td> 1,38</td>

<td>hsa-miR-548b</td><td> 1988</td><td> 1,92</td><td> -0,01</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-609</td><td> 1899</td><td> 2,55</td><td> 2,58</td><td> 3,6</td><td> 3,54</td>

<td>hsa-miR-377</td><td> 1929</td><td> 1,74</td><td> -0,01</td><td> 2,6</td><td> 0,58</td>

<td>hsa-miR-433</td><td> 1937</td><td> 2,71</td><td> 2,19</td><td> 3,74</td><td> 1,08</td>

<td>hsa-miR-500</td><td> 1957</td><td> 4,67</td><td> 4,88</td><td> 6,34</td><td> 5, 8</td>

<td>hsa-miR-652</td><td> 1961</td><td> 6,26</td><td> 6, 6</td><td> 5, 52</td><td> 5,05</td>

<td>hsa-miR-518c</td><td> 1981</td><td> 0, 92</td><td> 1,81</td><td> 3, 68</td><td> 1,38</td>

<td>hsa-miR-561</td><td> 1985</td><td> -0,58</td><td> 2,49</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-551a</td><td> 2018</td><td> 3,77</td><td> 3,27</td><td> 4,06</td><td> 3,91</td>

<td>hsa-miR-525</td><td> 2034</td><td> -0,58</td><td> 2,06</td><td> 3,1</td><td> 0,58</td>

<td>hsa-miR-570</td><td> 2054</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-617</td><td> 2062</td><td> 2</td><td> 2,49</td><td> 0,51</td><td> 2,91</td>

<td>hsa-miR-556</td><td> 2070</td><td> -0, 58</td><td> 1,81</td><td> 1,31</td><td> 1,08</td>

<td>hsa-miR-551b</td><td> 2074</td><td> 1,37</td><td> 1,79</td><td> 3,97</td><td> 3,38</td>

<td>hsa-miR-424</td><td> 1993</td><td> 5, 87</td><td>ç. o / xj tu 4</td><td>A CA hi _f “3</td><td>A 7 0 1 _z / -/</td>

<td>hsa-miR-612</td><td> 1997</td><td> 2,87</td><td> 0,29</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-130a</td><td> 2005</td><td> 8,07</td><td> 8,66</td><td> 9, 08</td><td> 9,2</td>

<td>hsa-miR-429</td><td> 2134</td><td> 5, 68</td><td> 5,39</td><td> 3,89</td><td> 4,83</td>

<td>hsa-miR-365</td><td> 2138</td><td> 8,7</td><td> 8,73</td><td> 7,51</td><td> 7,31</td>

<td>hsa-miR-29a</td><td> 2154</td><td> 13,45</td><td> 13,83</td><td> 12,21</td><td> 12,27</td>

<td>hsa-miR-503</td><td> 2162</td><td> 5, 44</td><td> 6,25</td><td> 1,31</td><td> 4,39</td>

<td>hsa-miR-624</td><td> 2166</td><td> -0,58</td><td> 1,99</td><td> 0,51</td><td> 3,39</td>

<td>hsa-miR-550</td><td> 2097</td><td> 4,34</td><td> 4,26</td><td> 3,89</td><td> 2,58</td>

<td>hsa-miR-581</td><td> 2227</td><td> 2,32</td><td> 1,65</td><td> 2,31</td><td> 0,58</td>

<td>hsa-miR-422a</td><td> 2263</td><td> 8,33</td><td> 8,48</td><td> 9,59</td><td> 9,25</td>

<td>hsa-miR-449</td><td> 2267</td><td> 2,91</td><td> 2,48</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-585</td><td> 2271</td><td> 3, 58</td><td> 3,74</td><td> 4,51</td><td> 4,12</td>

<td>hsa-miR-92b</td><td> 2182</td><td> 7,86</td><td> 8,04</td><td> 7,75</td><td> 7,13</td>

<td>hsa-miR-629</td><td> 2316</td><td> 6,12</td><td> 5, 93</td><td> 6, 82</td><td> 7,03</td>

<td>hsa-miR-580</td><td> 2320</td><td> -0,58</td><td> 1,49</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-448</td><td> 2324</td><td> 1,74</td><td> -0,51</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-592</td><td> 2332</td><td> 0,21</td><td> 2,95</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-671</td><td> 2839</td><td> 4,15</td><td> 3,98</td><td> 4,21</td><td> 0,58</td>

<td>hsa-miR-767-3p</td><td> 2863</td><td> 1,42</td><td> 2,3</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-608</td><td> 2279</td><td> 3,74</td><td> 1,29</td><td> 2,47</td><td> 0,58</td>

<td>hsa-miR-210</td><td> 2291</td><td> 9, 13</td><td> 8,6</td><td> 8,39</td><td> 7,96</td>

<td>hsa-miR-26a</td><td> 2299</td><td> 12,6</td><td> 12,61</td><td> 12,27</td><td> 12,73</td>

<td>hsa-miR-4 93-5p</td><td> 2329</td><td> 2</td><td> 2,65</td><td> 2,17</td><td> 1,08</td>

<td>hsa-miR-202 *</td><td> 2337</td><td> 2,55</td><td> 2,08</td><td> 3,32</td><td> 1,88</td>

<td>hsa-miR-454-5p</td><td> 2840</td><td> 11,51</td><td> 11,6</td><td> 12,87</td><td> 13,07</td>

<td>hsa-miR-770-5p</td><td> 2844</td><td> 2,24</td><td> -0,01</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-769-3p</td><td> 2865</td><td> 3,8</td><td> 3,74</td><td> 3,01</td><td> 2,88</td>

<td>hsa-miR-758</td><td> 2869</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-765</td><td> 2864</td><td> 5,35</td><td> 5, 13</td><td> 6, 08</td><td> 5,77</td>

<td>hsa-miR-301</td><td> 1103</td><td> 3, 96</td><td> 4,08</td><td> 4,04</td><td> 4,68</td>

<td>hsa-miR-191</td><td> 1017</td><td> 9,84</td><td> 10,87</td><td> 11,01</td><td> 10,9</td>

<td>hsa-miR-93</td><td> 1029</td><td> 9, 68</td><td> 9, 58</td><td> 10,93</td><td> 9,75</td>

<td>hsa-let-7f</td><td> 1033</td><td> 11,59</td><td> 12,43</td><td> 11,79</td><td> 11,94</td>

<td>hsa-miR-373</td><td> 1037</td><td> 3,54</td><td> 2,4</td><td> 4,51</td><td> 3,08</td>

<td>hsa-miR-200b</td><td> 1042</td><td> 10,9</td><td> 11,07</td><td> 10,17</td><td> 9,02</td>

<td>hsa-miR-100</td><td> 1064</td><td> 7,25</td><td> 6, 69</td><td> 7,81</td><td> 5,54</td>

<td>hsa-miR-324-3p</td><td> 1082</td><td> 5, 67</td><td> 4,71</td><td> 5, 64</td><td> 3, 99</td>

<td>hsa-miR-34b</td><td> 1096</td><td> 3,27</td><td> 3,49</td><td> 3,83</td><td> 4,54</td>

<td>hsa-miR-324-5p</td><td> 1115</td><td> 3,84</td><td> 2,29</td><td> 4,16</td><td> 4,49</td>

<td>hsa-miR-199a *</td><td> 1124</td><td> 1,82</td><td> 2,24</td><td> 1,01</td><td> 4,17</td>

<td>hsa-miR-103</td><td> 1164</td><td> 11,27</td><td> 10, 65</td><td> 11,3</td><td> 9, 18</td>

<td>hsa-miR-220</td><td> 1173</td><td> 3,67</td><td> 3,04</td><td> 4,26</td><td> 3, 99</td>

<td>hsa-miR-151</td><td> 1199</td><td> 9,73</td><td> 9, 47</td><td> 10,22</td><td> 10,45</td>

<td>hsa-miR-18 6</td><td> 1141</td><td> 4,72</td><td> 4,93</td><td> 3,86</td><td> 4,49</td>

<td>hsa-miR-128b</td><td> 1153</td><td> 6,29</td><td> 6,26</td><td> 6,7</td><td> 6,1</td>

<td>hsa-miR-130b</td><td> 1165</td><td> 7,72</td><td> 6, 96</td><td> 7,99</td><td> 6,49</td>

<td>hsa-miR-338</td><td> 1174</td><td> 2,42</td><td> 2,66</td><td> 2,67</td><td> 2,91</td>

<td>hsa-miR-199b</td><td> 1178</td><td> 1,98</td><td> -0,01</td><td> 3, 67</td><td> 3,46</td>

<td>hsa-miR-125b</td><td> 1182</td><td> 9,34</td><td> 8,81</td><td> 9,86</td><td> 8,11</td>

<td>hsa-miR-122a</td><td> 1243</td><td> 5,11</td><td> 3,49</td><td> 4,97</td><td> 4,71</td>

<td>hsa-miR-30d</td><td> 1251</td><td> 11,72</td><td> 11,93</td><td> 11,32</td><td> 11,69</td>

<td>hsa-miR-203</td><td>ί '-ί rr \ 1Z OR</td><td>Ί / IO i / * yay,</td><td>OQ</td><td> ,—1</td><td> 9,56</td>

<td>hsa-let-7c</td><td> 1268</td><td> 11, 91</td><td> 12,72</td><td> 13,09</td><td> 12,47</td>

<td>hsa-miR-216</td><td> 1294</td><td> 2</td><td> 2,45</td><td> 2,71</td><td> 3,38</td>

<td>hsa-miR-15b</td><td> 1313</td><td> 11,75</td><td> 12,27</td><td> 12,66</td><td> 12,77</td>

<td>hsa-miR-192</td><td> 1205</td><td> 7,05</td><td> 8,48</td><td> 6</td><td> 6,14</td>

<td>hsa-miR-133a</td><td> 1215</td><td> 3,27</td><td> 3,07</td><td> 3,82</td><td> 4,11</td>

<td>hsa-miR-12 6</td><td> 1380</td><td> 6, 42</td><td> 6, 42</td><td> 5, 94</td><td> 7,51</td>

<td>hsa-miR-326</td><td> 1393</td><td> 3,32</td><td> 0,29</td><td> 0,51</td><td> 3,17</td>

<td>hsa-miR-98</td><td> 1423</td><td> 6, 58</td><td> 7,21</td><td> 6,9</td><td> 7,33</td>

<td>hsa-let-7g</td><td> 1432</td><td> 10,8</td><td> 11,21</td><td> 10,01</td><td> 10,06</td>

<td>hsa-miR-190</td><td> 1437</td><td> 3,16</td><td> 3, 57</td><td> 4,02</td><td> 4,29</td>

<td>hsa-miR-189</td><td> 1442</td><td> 2,59</td><td> 2,79</td><td> 2,92</td><td> 3,38</td>

<td>hsa-miR-137</td><td> 1339</td><td> 2,66</td><td> 3,06</td><td> 4,36</td><td> 3,88</td>

<td>hsa-miR-105</td><td> 1345</td><td> 2,37</td><td> 2,48</td><td> 4,32</td><td> 3,17</td>

<td>hsa-miR-96</td><td> 1507</td><td> 4,66</td><td> 4,17</td><td> 4,58</td><td> 4,58</td>

<td>hsa-miR-519e</td><td> 1767</td><td> -0,08</td><td> 2,3</td><td> 2,83</td><td> 1,08</td>

<td>hsa-miR-520a</td><td> 1771</td><td> 1,42</td><td> 1,99</td><td> 3,1</td><td> 0,58</td>

<td>hsa-miR-451</td><td> 1783</td><td> 1</td><td> -0,51</td><td> 3,32</td><td> 2,58</td>

<td>hsa-miR-523</td><td> 1787</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-299-5p</td><td> 1458</td><td> 1,74</td><td> 2,06</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-95</td><td> 1482</td><td> 3,94</td><td> 3,78</td><td> 2,71</td><td> 3,08</td>

<td>hsa-miR-593</td><td> 1832</td><td> 4,92</td><td> 1,08</td><td> 4,1</td><td> 2,91</td>

<td>hsa-miR-57 4</td><td> 1840</td><td> 11,34</td><td> 9, 36</td><td> 11,12</td><td> 9, 45</td>

<td>hsa-miR-202</td><td> 1864</td><td> 2,59</td><td> 1,9</td><td> 2, 97</td><td> 3, 67</td>

<td>hsa-miR-519b</td><td> 1799</td><td> -0,08</td><td> 3,49</td><td> 3, 97</td><td> 3,83</td>

<td>hsa-miR-520d</td><td> 1803</td><td> 2,58</td><td> 3,56</td><td> 3, 89</td><td> 4,36</td>

<td>hsa-miR-622</td><td> 1829</td><td> 1,42</td><td> 2,66</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-483</td><td> 1845</td><td> 8,06</td><td> 3,91</td><td> 7,36</td><td> 4,34</td>

<td>hsa-miR-383</td><td> 1865</td><td> 2,17</td><td> 1,95</td><td> 0,51</td><td> 1,88</td>

<td>hsa-miR-29b</td><td> 1869</td><td> 6, 67</td><td> 6,05</td><td> 5,9</td><td> 6, 65</td>

<td>hsa-miR-613</td><td> 1881</td><td> 2,42</td><td> -0,01</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-453</td><td> 1904</td><td> 3,93</td><td> 3,59</td><td> 4,76</td><td> 4,17</td>

<td>hsa-miR-23b</td><td> 1930</td><td> 13,2</td><td> 13,81</td><td> 12,99</td><td> 13,61</td>

<td>hsa-miR-501</td><td> 1942</td><td> 2,87</td><td> 2,72</td><td> 1, 31</td><td> 3,25</td>

<td>hsa-miR-517c</td><td> 1946</td><td> 3,01</td><td> 3,02</td><td> 4,36</td><td> 4,39</td>

<td>hsa-miR-625</td><td> 1897</td><td> 6, 54</td><td> 7,16</td><td> 5, 67</td><td> 5, 65</td>

<td>hsa-miR-630</td><td> 1905</td><td> 2,74</td><td> 1,49</td><td> 2,92</td><td> 3,49</td>

<td>hsa-miR-644</td><td> 1913</td><td> 0,21</td><td> 2,29</td><td> 2,51</td><td> 2,91</td>

<td>hsa-miR-527</td><td> 2039</td><td> 3, 42</td><td> 1,87</td><td> 2, 51</td><td> 1, 08</td>

<td>hsa-miR-589</td><td> 2055</td><td> -0,08</td><td> -0,51</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-508</td><td> 2071</td><td> 2,81</td><td> 2,79</td><td> 3,21</td><td> 4,04</td>

<td>hsa-miR-449b</td><td> 2083</td><td> 2,41</td><td> 2,48</td><td> 3, 97</td><td> 2,58</td>

<td>hsa-miR-603</td><td> 1990</td><td> 4,74</td><td> 2,84</td><td> 3,76</td><td> 0,58</td>

<td>hsa-miR-200c</td><td> 2131</td><td> 4,25</td><td> 3,75</td><td> 13,3</td><td> 13,7</td>

<td>hsa-miR-29c</td><td> 2155</td><td> 2,75</td><td> 3,24</td><td> 3, 6</td><td> 4,2</td>

<td>hsa-miR-411</td><td> 2167</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-455</td><td> 2179</td><td> 2,87</td><td> 2,52</td><td> 2,6</td><td> 0,58</td>

<td>hsa-miR-378</td><td> 2208</td><td> 2,24</td><td> 2,3</td><td> 3, 6</td><td> 2,58</td>

<td>hsa-miR-638</td><td> 2212</td><td> 7,37</td><td> 6,34</td><td> 8,56</td><td> 7,57</td>

<td>hsa-miR-518f *</td><td> 2220</td><td> -0,58</td><td> -0,51</td><td> 3, 17</td><td> 3,67</td>

<td>hsa-let-7i</td><td> 2244</td><td> 12,8</td><td> 13,03</td><td> 10,86</td><td> 10,79</td>

<td>hsa-miR-422b</td><td> 2264</td><td> 9, 17</td><td> 8,72</td><td> 10,51</td><td> 10,23</td>

<td>hsa-miR-193b</td><td> 2268</td><td> 9, 68</td><td> 8,44</td><td> 8,63</td><td> 7,54</td>

<td>hsa-miR-491</td><td> 2272</td><td> 1,74</td><td> 0,79</td><td> 2,81</td><td> 0,58</td>

<td>hsa-miR-484</td><td> 2191</td><td> 8,32</td><td> 7,81</td><td> 8,29</td><td> 7,72</td>

<td>hsa-miR-623</td><td> 2203</td><td> 1,74</td><td> 2,45</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-486</td><td> 2209</td><td> 3,86</td><td> 3, 3</td><td> 4,2</td><td> 4,6</td>

<td>hsa-miR-639</td><td> 2213</td><td> 1,87</td><td> 1,49</td><td> 2,31</td><td> 1,38</td>

<td>hsa-miR-517a hsamiR-517b</td><td> 2217</td><td> 2,11</td><td> 2,56</td><td> 3, 87</td><td> 3,28</td>

<td>hsa-miR-645</td><td> 2221</td><td> 3,12</td><td> 1,29</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-146b</td><td> 2237</td><td> 5,56</td><td> 5,29</td><td> 4,21</td><td> 5,59</td>

<td>hsa-miR-571</td><td> 2249</td><td> 3,33</td><td> 2,99</td><td> 4,1</td><td> 2,91</td>

<td>hsa-miR-191 *</td><td> 2257</td><td> 2,42</td><td> 2,95</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-7</td><td> 2261</td><td> 2,44</td><td> 3,02</td><td> 2,51</td><td> 3,54</td>

<td>hsa-miR-647</td><td> 2269</td><td> 4,95</td><td> 4,27</td><td> 5, 5</td><td> 6,01</td>

<td>hsa-miR-637</td><td> 2273</td><td> 4,65</td><td> 2,84</td><td> 4,9</td><td> 4,17</td>

<td>hsa-miR-30b</td><td> 2280</td><td> 9, 94</td><td> 9, 87</td><td> 9,86</td><td> 9, 66</td>

<td>hsa-miR-431</td><td> 2288</td><td> 1,74</td><td> -0,01</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-452</td><td> 2292</td><td> 4,68</td><td> 5,15</td><td> 5, 14</td><td> 5,85</td>

<td>hsa-miR-361</td><td> 2296</td><td> 10,36</td><td> 11,32</td><td> 10,53</td><td> 10,83</td>

<td>hsa-miR-57 6</td><td> 2314</td><td> 1,87</td><td> -0,51</td><td> 2, 83</td><td> 0,58</td>

<td>hsa-miR-432</td><td> 2326</td><td> 3,74</td><td> 3, 47</td><td> 3, 51</td><td> 2,58</td>

<td>hsa-miR-375</td><td> 2342</td><td> 3,42</td><td> 2,15</td><td> 0, 51</td><td> 3,75</td>

<td>hsa-miR-766</td><td> 2841</td><td> 9, 66</td><td> 6, 37</td><td> 8,18</td><td> 7,59</td>

<td>hsa-miR-7 68-3p</td><td> 2845</td><td> 9,89</td><td> 9, 61</td><td> 9,2</td><td> 9, 48</td>

<td>hsa-miR-7 69-5p</td><td> 2861</td><td> 4,03</td><td> 4,07</td><td> 4, 47</td><td> 3,46</td>

<td>hsa-miR-513</td><td> 2301</td><td> 3,8</td><td> 2,56</td><td> 3, 97</td><td> 4,38</td>

<td>hsa-miR-362</td><td> 2017</td><td> 2,93</td><td> 4,53</td><td> 4,88</td><td> 4,38</td>

<td>hsa-miR-565</td><td> 2045</td><td> 7,04</td><td> 4,89</td><td> 5,13</td><td> 6, 45</td>

<td>hsa-miR-30e-3p</td><td> 2053</td><td> 8,97</td><td> 9,4</td><td> 7,84</td><td> 7,61</td>

<td>hsa-miR-320</td><td> 1005</td><td> 12,75</td><td> 13,28</td><td> 13, 11</td><td> 13, 09</td>

<td>hsa-miR-132</td><td> 1014</td><td> 4,94</td><td> 6, 57</td><td> 6,22</td><td> 7,04</td>

<td>hsa-miR-193a</td><td> 1018</td><td> 4,56</td><td> 4,32</td><td> 3, 66</td><td> 4,58</td>

<td>hsa-miR-22</td><td> 1022</td><td> 8,71</td><td> 8,95</td><td> 8, 69</td><td> 8,79</td>

<td>hsa-miR-224</td><td> 1026</td><td> 6, 69</td><td> 7,1</td><td> 6,4</td><td> 6, 96</td>

<td>hsa-let-7a</td><td> 1030</td><td> 13,37</td><td> 14,07</td><td> 14,63</td><td> 14,91</td>

<td>hsa-miR-302d</td><td> 1034</td><td> 2,32</td><td> 2,74</td><td> 3,76</td><td> 3,28</td>

<td>hsa-miR-369-3p</td><td> 1038</td><td> 2,72</td><td> 2,38</td><td> 4,68</td><td> 3,83</td>

<td>hsa-miR-106a</td><td> 1006</td><td> 12,01</td><td> 12,48</td><td> 12,09</td><td> 12,36</td>

<td>hsa-miR-181c</td><td> 1015</td><td> 5, 67</td><td> 6,09</td><td> 4,64</td><td> 4,27</td>

<td>hsa-miR-17-5p</td><td> 1031</td><td> 11,57</td><td> 11,85</td><td> 11,34</td><td> 11,83</td>

<td>hsa-miR-302b</td><td> 1035</td><td> -0,08</td><td> 2,66</td><td> 3,26</td><td> 4,04</td>

<td>hsa-miR-19b</td><td> 1039</td><td> 10,14</td><td> 10,07</td><td> 11,3</td><td> 11,47</td>

<td>hsa-miR-24</td><td> 1044</td><td> 12,91</td><td> 13,2</td><td> 13,13</td><td> 13, 4</td>

<td>hsa-miR-17-3p</td><td> 1079</td><td> 4,95</td><td> 5,02</td><td> 4,83</td><td> 5, 34</td>

<td>hsa-miR-221</td><td> 1088</td><td> 13,67</td><td> 13,73</td><td> 12,88</td><td> 12,76</td>

<td>hsa-miR-335</td><td> 1146</td><td> -0,58</td><td> -0,51</td><td> 6, 66</td><td> 7, 68</td>

<td>hsa-miR-199a</td><td> 1167</td><td> 2,31</td><td> -0,51</td><td> 0,51</td><td> 3,17</td>

<td>hsa-miR-126 *</td><td> 1171</td><td> 3,12</td><td> 1,95</td><td> 3,68</td><td> 3,15</td>

<td>hsa-miR-337</td><td> 1175</td><td> 2,22</td><td> -0,51</td><td> 3,97</td><td> 2,91</td>

<td>hsa-miR-181a *</td><td> 1179</td><td> 5, 67</td><td> 5, 34</td><td> 5,91</td><td> 5,76</td>

<td>hsa-miR-331</td><td> 1183</td><td> 6,46</td><td> 5,25</td><td> 5,55</td><td> 4,95</td>

<td>hsa-miR-340</td><td> 1187</td><td> 2,96</td><td> 2,99</td><td> 3,86</td><td> 4,17</td>

<td>hsa-miR-188</td><td> 1116</td><td> 3,94</td><td> 3,31</td><td> 3,8 6</td><td> 4,39</td>

<td>hsa-miR-9</td><td> 1231</td><td> 2,96</td><td> 3,25</td><td> 4</td><td> 4,53</td>

<td>hsa-miR-34a</td><td> 1235</td><td> 6,95</td><td> 6, 56</td><td> 7,17</td><td> 7,33</td>

<td>hsa-miR-30c</td><td> 1252</td><td> 13,78</td><td> 13,97</td><td> 12,46</td><td> 12,24</td>

<td>hsa-miR-19a</td><td> 1271</td><td> 5, 93</td><td> 5, 76</td><td> 8,01</td><td> 8,36</td>

<td>hsa-miR-371</td><td> 1276</td><td> 3,67</td><td> 2,19</td><td> 3,36</td><td> 3, 38</td>

<td>hsa-miR-lOb</td><td> 1301</td><td> 6, 91</td><td> 7,36</td><td> 7,73</td><td> 8,03</td>

<td>hsa-miR-21</td><td> 1315</td><td> 13,13</td><td> 13,2</td><td> 12,28</td><td> 12,88</td>

<td>hsa-miR-217</td><td> 1206</td><td> 2,53</td><td> 2,49</td><td> 0,51</td><td> 3,57</td>

<td>hsa-miR-302b *</td><td> 1210</td><td> 1,87</td><td> 2,49</td><td> 2,51</td><td> 2, 99</td>

<td>hsa-miR-135a</td><td> 1216</td><td> 2,41</td><td> 3, 62</td><td> 3,47</td><td> 3,89</td>

<td>hsa-miR-148a</td><td> 1361</td><td> 3</td><td> 1,45</td><td> 6, 87</td><td> 7,35</td>

<td>hsa-miR-339</td><td> 1366</td><td> 4,85</td><td> 4,26</td><td> 5, 12</td><td> 5,2</td>

<td>hsa-miR-187</td><td> 1381</td><td> 3,69</td><td> 2,4</td><td> 4,21</td><td> 3,75</td>

<td>hsa-miR-346</td><td> 1390</td><td> 5,77</td><td> 3,2</td><td> 4,09</td><td> 4,87</td>

<td>hsa-miR-146a</td><td> 1409</td><td> 9,7</td><td> 9,88</td><td> 7,17</td><td> 7,56</td>

<td>hsa-miR-143</td><td> 1415</td><td> -0,58</td><td> -0,51</td><td> 2,51</td><td> 3,75</td>

<td>hsa-miR-219</td><td> 1426</td><td> 2</td><td> 1,81</td><td> 3,32</td><td> 4,04</td>

<td>hsa-miR-185</td><td> 1451</td><td> 8,4</td><td> 8,73</td><td> 9,33</td><td> 9,46</td>

<td>hsa-miR-328</td><td> 1455</td><td> 7,15</td><td> 4,5</td><td> 4,92</td><td> 4,33</td>

<td>hsa-miR-196b</td><td> 1321</td><td> 4,65</td><td> 4,44</td><td> 5,08</td><td> 5, 68</td>

<td>hsa-miR-204</td><td> 1489</td><td> 0,71</td><td> 2,49</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-133b</td><td> 1498</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-129</td><td> 1512</td><td> 6,33</td><td> 6, 08</td><td> 7,2</td><td> 8,02</td>

<td>hsa-miR-649</td><td> 1756</td><td> 3,32</td><td> 2,93</td><td> 3, 17</td><td> 2,17</td>

<td>hsa-miR-522</td><td> 1776</td><td> 2,87</td><td> 3,4</td><td> 5, 74</td><td> 5, 87</td>

<td>hsa-miR-618</td><td> 1788</td><td> 2,22</td><td> 1,65</td><td> 0,51</td><td> 1,08</td>

<td>hsa-miR-30a-5p</td><td> 1460</td><td> 12,45</td><td> 12,55</td><td> 11,09</td><td> 11,04</td>

<td>hsa-miR-27a</td><td> 1485</td><td> 11,64</td><td> 11,67</td><td> 11,97</td><td> 12,27</td>

<td>hsa-miR-30a-3p</td><td> 1505</td><td> 12,22</td><td> 12,57</td><td> 10</td><td> 10,48</td>

<td>hsa-miR-4 94</td><td> 1753</td><td> 4,47</td><td> 3, 87</td><td> 6, 12</td><td> 5, 48</td>

<td>hsa-miR-20b</td><td> 1769</td><td> 10,41</td><td> 10,8</td><td> 10,92</td><td> 11,2</td>

<td>hsa-miR-521</td><td> 1785</td><td> 3,42</td><td> 0,49</td><td> 3,31</td><td> 3,75</td>

<td>hsa-miR-363</td><td> 1822</td><td> -0, 58</td><td> -0, 51</td><td> 3,32</td><td> 1,08</td>

<td>hsa-miR-181b</td><td> 1830</td><td> 11,53</td><td> 11,96</td><td> 10,84</td><td> 11,02</td>

<td>hsa-miR-18a *</td><td> 1850</td><td> 4,52</td><td> 2,99</td><td> 4,97</td><td> 3,83</td>

<td>hsa-miR-423</td><td> 1874</td><td> 8,9</td><td> 8,85</td><td> 9, 09</td><td> 8,46</td>

<td>hsa-miR-595</td><td> 1805</td><td> 9,11</td><td> 6, 55</td><td> 8,47</td><td> 6,49</td>

<td>hsa-miR-487b</td><td> 1817</td><td> 4,65</td><td> 4,3</td><td> 5,22</td><td> 5, 53</td>

<td>hsa-miR-425-3p</td><td> 1943</td><td> 4,14</td><td> 4,02</td><td> 3, 39</td><td> 3, 96</td>

<td>hsa-miR-594</td><td> 1951</td><td> 10,94</td><td> 10,48</td><td> 11,55</td><td> 11,22</td>

<td>hsa-miR-532</td><td> 1959</td><td> 5,87</td><td> 5, 79</td><td> 6, 62</td><td> 6, 14</td>

<td>hsa-miR-544</td><td> 1971</td><td> 1,08</td><td> 2,49</td><td> 1,01</td><td> 2,91</td>

<td>hsa-miR-512-3p</td><td> 1910</td><td> 2,56</td><td> 2,74</td><td> 4,41</td><td> 3,83</td>

<td>hsa-miR-526a</td><td> 2036</td><td> -0,58</td><td> -0,51</td><td> 5,78</td><td> 5,87</td>

<td>hsa-miR-619</td><td> 2044</td><td> 2,01</td><td> 1,49</td><td> 1,01</td><td> 4,08</td>

<td>hsa-miR-578</td><td> 2048</td><td> 3,54</td><td> 2,79</td><td> 3,17</td><td> 2,38</td>

<td>hsa-miR-4 92</td><td> 2060</td><td> -0, 08</td><td> 1,49</td><td> 2,71</td><td> 2,67</td>

<td>hsa-miR-590</td><td> 2064</td><td> 3,27</td><td> 3,4</td><td> 5,51</td><td> 5,08</td>

<td>hsa-miR-515-3p</td><td> 2068</td><td> 1,74</td><td> 2,88</td><td> 3,51</td><td> 1,08</td>

<td>hsa-miR-539</td><td> 2080</td><td> 2,74</td><td> 1,81</td><td> 2,51</td><td> 4,28</td>

<td>hsa-miR-497</td><td> 1995</td><td> 3,05</td><td> 3,11</td><td> 3,26</td><td> 0,58</td>

<td>hsa-miR-152</td><td> 2007</td><td> 7,72</td><td> 8,44</td><td> 6, 59</td><td> 7,2</td>

<td>hsa-miR-181d</td><td> 2011</td><td> 8,56</td><td> 8,9</td><td> 7,83</td><td> 7,49</td>

<td>hsa-miR-660</td><td> 2144</td><td> 5, 3</td><td> 5,36</td><td> 6, 62</td><td>co</td>

<td>hsa-miR-584</td><td> 2176</td><td> 10,1</td><td> 10, 43</td><td> 7,6</td><td> 7,99</td>

<td>hsa-miR-511</td><td> 2109</td><td> 2,59</td><td> -0,01</td><td> 2,83</td><td> 2,91</td>

<td>hsa-miR-141</td><td> 2117</td><td> -0,58</td><td> -0,51</td><td> 7,91</td><td> 8,21</td>

<td>hsa-miR-18b</td><td> 2125</td><td> 5,18</td><td> 5, 41</td><td> 6, 65</td><td> 6, 98</td>

<td>hsa-miR-582</td><td> 2141</td><td> -0,58</td><td> -0,51</td><td> 4,9</td><td> 4,87</td>

<td>hsa-miR-58 6</td><td> 2173</td><td> 2,11</td><td> 1,49</td><td> 2,47</td><td> 1,08</td>

<td>hsa-miR-505</td><td> 2184</td><td> 5, 06</td><td> 5,45</td><td> 4,16</td><td> 4,58</td>

<td>hsa-miR-642</td><td> 2200</td><td> 4,22</td><td> 1,15</td><td> 2,42</td><td> 0,58</td>

<td>hsa-miR-628</td><td> 2222</td><td> 3,59</td><td> 2,19</td><td> 2,17</td><td> 3,83</td>

<td>hsa-miR-425-5p</td><td> 2234</td><td> 8,86</td><td> 9,29</td><td> 9,01</td><td> 8,92</td>

<td>hsa-miR-661</td><td> 2274</td><td> 2,42</td><td> 1,81</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-421</td><td> 2185</td><td> 4,06</td><td> 5, 49</td><td> 6, 41</td><td> 6,43</td>

<td>hsa-miR-27b</td><td> 2303</td><td> 10,82</td><td> 11,2</td><td> 11,39</td><td> 11,55</td>

<td>hsa-miR-651</td><td> 2335</td><td> 1,71</td><td> 1,69</td><td> 3,39</td><td> 2,91</td>

<td>hsa-miR-557</td><td> 2339</td><td> 3,37</td><td> 2,49</td><td> 3,51</td><td> 0,58</td>

<td>hsa-miR-801</td><td> 2846</td><td> 5, 97</td><td> 3,08</td><td> 4,59</td><td> 1,88</td>

<td>hsa-miR-768-5p</td><td> 2854</td><td> 8,68</td><td> 8,01</td><td> 8,5</td><td> 8,38</td>

<td>hsa-miR-454-3p</td><td> 2858</td><td> 3</td><td> 3,37</td><td> 4,32</td><td> 3,78</td>

<td>hsa-miR-498</td><td> 2298</td><td> 2,87</td><td> -0,51</td><td> 0,51</td><td> 2,67</td>

<td>hsa-miR-148b</td><td> 1362</td><td> 6, 83</td><td> 6,76</td><td> 6, 82</td><td> 6, 69</td>

<td>hsa-miR-194</td><td> 1416</td><td> 8,57</td><td> 8,28</td><td> 4,64</td><td> 5,81</td>

<td>hsa-let-7e</td><td> 1421</td><td> 7,42</td><td> 9, 18</td><td> 8,74</td><td> 9, 52</td>

<td>hsa-miR-345</td><td> 1444</td><td> 4,63</td><td> 4, 62</td><td> 3, 68</td><td> 3,17</td>

<td>hsa-miR-155</td><td> 1476</td><td> 8,21</td><td> 9,31</td><td> 4,32</td><td> 6,17</td>

<td>hsa-miR-374</td><td> 1480</td><td> 1,42</td><td> 1,79</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-26b</td><td> 1484</td><td> 9, 52</td><td> 10</td><td> 9, 72</td><td> 10,43</td>

* The raw data were subtracted from the background, Log2 transformed and normalized. The intensity of each oligo probe is based on the average of duplicated spots. The normalized limit is calculated based on log2 (5 * standard deviation of the 5 non-spot background + negative control probe signal with trim mean method)

TABLE 2

ASSOCIATION OF MYRNA WITH PERIPHERAL EXOSOMES

BLOOD DERIVED TUMOR COMPARED WITH MYRNA ISOLATED FROM YOURS

MATCHING TUMORS

Association of microRNA with peripheral blood-derived tumor exosomes compared with microRNA isolated from their corresponding tumors.

<td>Elevated in cells</td><td>Equal between cells and exosomes</td><td>Elevated in exosomes</td>

<td>miR-218, miR-196a, miR-195, miR-15a, miR-519d, miR-382, miR-503, miR-34b, miR-520d, miR-29c, miR-135a, miR-155</td><td>miR-296, miR-20a, miR-28, miR-302a, miR-99a, miR99b, miR-10a, let-7a, let7b, let-7c, let-7d, let7f, let-7g, let-7i, miR138, miR-23a, miR-183, miR-25, miR-107, miR-181a, miR-125a, miR-222, miR198, miR-16, miR-200a, miR-18a, miR-101, miR- 136, miR-31, miR-106b, miR-92, miR-342, miR-128a, miR182, miR-663, miR-502, miR-500, miR-652, miR-424, miR-130a, miR- 429, miR365, miR-29a, miR-550, miR-422a, miR-585, miR92b, miR-629, miR-671, miR-210, miR-26a, miR-4545p, miR-769-3p, mi- 765, miR-301, miR-191, miR-93, miR-200b, miR-100, miR324-5p, miR-220, miR-151, miR-186, miR-128b, miR130b, miR-125b, miR-122a, miR-30d, miR-203, miR-15b, miR-192, miR-133a, miR126, miR-98, miR-190, miR137, miR-105, miR-96, miR95, miR-519b, miR-29b, miR-453, miR-23b, miR517c, miR-625, miR-200c, miR-193a, miR-22, miR-224, miR-369-3p, miR-106a, miR181c, miR-17-5p, miR-19b, miR-24, miR- 17-3p, miR221, miR-335, miR-126, miR-181a, miR-331, miR188, miR-9, miR-34a, miR30c, miR-19a, miR-371, miR-lOb, miR-21, miR-148a, miR-339, miR-187, miR-346, miR-146a, miR-185, miR328, miR-196b, miR-129,</td><td>miR-214, miR-140, miR-147, miR135b, miR-205, miR-150, miR-149, miR-370, miR-206, miR-197, miR-634, miR-485-5p, miR612, miR-608, miR-202, miR-373, miR-324-3p, miR103, miR-593, miR-574, miR-483, miR-527, miR-603, miR-649, miR-18a, miR- 595, miR193b, miR-642, miR-557, miR-801, let-7e</td>

<td>Elevated in cells</td><td>Equal between cells and exosomes</td><td>Elevated in exosomes</td>

<td></td><td>miR-522, miR-30a-5p, miR27a, miR-30a-3p, miR-494, miR-20b, miR-521, miR181b, miR-423, miR-487b, miR-425-3p, miR-594, miR532, miR-512-3p, miR-526a, miR-578, miR-638, miR422b, miR-484, miR-486, miR-645, miR-146b, miR571, miR-647, miR-637, miR- 30b, miR-452, miR-361, miR-432, miR-375, miR-766, miR-768-3p, miR-769-5p, miR-513, miR-362, miR-565, miR-30e- 3p, miR-320, miR590, miR-152, miR-181d, miR-660, miR-584, miR-141, miR-18b, miR-582, miR-505, miR-628, miR-425-5p, miR421, miR-27b, miR-768-5p, miR-454-3p, miR-148b, miR194, miR-345, miR-26b</td><td></td>

EXAMPLE 4

CORRELATION OF EXOSOMAL MYRNA WITH THE PRESENCE

DISEASE STAGE

Our previous comparisons between the tumor and the circulating exosomes were made in patients in advanced stage. To compare the associations of specific miRNAs with the presence of the disease in the various stages, the mean intensities of exosomal miRNAs were determined. The presence of the 8 diagnostic miRNAs among patients with stages I, II and III was not significantly different from the majority of these miRNAs (Figure 6).

The miR-200c and miR-214 were lower in patients in stage I, compared with stages II and III. However, in all cases, these miRNAs were significantly elevated from the levels detected in exosomes derived from benign disease. The fraction of small RNA could not be demonstrated in normal controls, and attempts to assess the presence of miRNAs were negative.

EXAMPLE 5

STABILITY OF EXOSOMAL MIRNA PROFILES

As the measurement of the circulating exosomal miRNA was demonstrated here as being diagnostic, the technical question of its stability was later verified. When miRNA profiles were made in serum samples stored for short periods at 4 ° C (up to 96 hours) and the intensities were compared (Figure 7A), no significant differences were observed in the 3 diagnostic miRNAs analyzed. When serum samples were stored at -70 ° C for longer periods of time, the intensities of these miRNAs in the microarrays were not significantly different (Figure 7B). These results indicate that the levels of these exosomal miRNA were stable and did not change significantly with storage.

DISCUSSION OF EXAMPLES 1-5

The profiling of microRNA expression can be used as a diagnostic tool for cancers that currently do not have reliable molecular markers, such as ovarian cancer. Although previous studies have indicated that miRNA signatures could serve as a diagnosis and prognostic markers for ovarian cancer, these data were based on their expressions in tissue samples. The present Examples provide data that demonstrate for the first time the association of miRNA with circulating tumor-derived exosomes. In previous studies, miRNAs have been shown to be expressed in an aberrant way in human ovarian cancers and the total miRNA expression could differentiate normal tissue from cancer tissue (Lorio et al., 2007). The study by Lu et al. (2005) demonstrated the use of miRNA signatures as an important advance in the diagnosis of cancer. This work indicated that miRNA-based identification of cancers was superior for the correct diagnosis of unknown primary cancers with respect to mRNA classification.

However, prior to the material currently revealed, it was not possible to use miRNA profiling in the absence of a biopsy mass.

Our original electron microscopic characterization of the exosomes indicated that they were hollow (that is, the absence of viral-like structures) (Taylor &amp; Black, 1986). As a result, our group, together with the others, focused on the external protein components of the exosomes and the biological consequences of exosome exposure. In the present Examples, however, we have surprisingly demonstrated, for the first time, the presence of small species of RNA associated with circulating tumor exosomes (Figure 4). This small RNA does not have the 18S and 28S associated with the cell's RNA. In addition, the microarray analysis presented in the present study demonstrated that at least part of the small RNA identified is miRNA.

The miRNA expression profiles of our ovarian tumor cells confirmed the miRNA aberrations indicated in previous studies. Analyzes of circulating tumor exosomes and tumor cells from the same patients showed that both were positive for 46% of the tested miRNAs (218/467). When the miRNA intensities normalized, most of these miRNAs were expressed at similar levels between cells and exosomes or increased within exosomes (175 were not significantly different and 31 were elevated within exosomes). Thus, the aberrantly expressed miRNAs, used to establish specific cancer signatures, appear in both the cellular and exosomal compartments of patients with ovarian cancer.

Our comparison of specific miRNAs, previously shown to be diagnostic, indicated a high degree of correlation between the tumor's miRNA and its corresponding exosomes (ranging between 0.71 and 0.90). This high correlation even includes miRNAs that appeared to be present in greater proportions in exosomes, as for miR-214. The uniform elevation of specific miRNAs in exosomes has led to the suggestion that the compartmentalization of miRNAs in exosomes, in at least some miRNAs, is an active (selective) process. This process could be mediated by components, such as nucleolin or nucleophosmin, which are aberrantly expressed in tumor exosomes.

As these results demonstrated that the exosomal miRNA profiling can be used as a substitute for tissue miRNA and the purpose of the scan would be to identify the disease in the early stages, the ability to detect exosomal circulating exosomal miRNA in the early stages of the disease was examined. The exosomal miRNA expressions of diagnostic miRNAs between patients with early and late stage ovarian cancers were not significantly different from most of these miRNAs (Figure 6). MiR-200c and miR-214 were lower in patients in stage I, compared to stages II and III;

however, in all cases, these miRNAs were significantly elevated in relation to the levels detected in exosomes derived from benign disease. The small RNA fraction could not be demonstrated in normal controls and attempts to assess the presence of miRNAs were negative. Thus, the absence of exosomes and / or small exosomal RNAs is associated with normal individuals, who do not have cancer, and the normal tissue mirroring exosomal miRNA seems to be associated with benign disease. The similarity between the stages of ovarian cancer is the likely result of the standardization of quantities of small exosomal starting RNAs and the normalization of the resulting arrangement data. Despite this standardization and normalization, the profiles obtained with exosomal miRNA from patients with benign disease remained distinct. These results demonstrate that analyzes of specific miRNAs associated with circulating exosomes can be applied to all stages of ovarian cancer and that benign and malignant diseases appear distinguishable based on the levels of the 8 specific miRNAs noted in the present.

The miRNA signatures of exosomes parallel to those of the miRNA expression profiles of the tumor cells of origin, indicating that the miRNA profiling can be done in the absence of tissue accurately reflect the tumor profile. We also observed that the tumor-derived exosomes of lung cancer patients contain miRNA similar to the corresponding tumor miRNA signatures (see Example 6). Circulating exosomes derived from the tumor can be isolated using tumor markers, such as EpCAM, followed by the analysis of miRNAs associated with the exosomes. Since this approach is non-invasive, as it does not need a biopsy mass, the exosomal miRNA profiling can be used as a scanning tool to detect many different cancers. How specific miRNAs associated with tumor tissues are identified as predictors of prognosis, including therapeutic resistance (such as let-7i, miR-16, miR-21 and miR-214) (Yang et al., 2008; Biower et al.,

2008), their presence in tumor exosomes can be evaluated to further define the usefulness of exosomal miRNA profiling as a prognostic indicator. The use of exosomal miRNA profiling can extend this approach to the scanning of asymptomatic individuals, as well as for monitoring the recurrence of the disease.

EXAMPLE 6

CORRELATION OF MYRNA WITH EXOSOMES FROM TUMORS OF

PERIPHERAL BLOOD DERIVED LUNG COMPARED TO miRNAs

ISOLATED FROM ITS CORRESPONDING LUNG TUMORS

In studies demonstrating diagnostic miRNA signatures of non-small cell lung carcinoma (NSCLC), specific miRNAs 20 were excessively expressed compared to normal lung tissue (miR-17-3p, miR-21, miR106a, m iR-146, miR- 155, miR-191, miR-192, miR-203, miR-205, miR210, miR-212 and miR-214). To correlate these findings with the material derived from the patients, miRNA fractions were isolated and profiled from circulating tumor exosomes and from the original tumor 25 using methods revealed above in the present and shown in Figure 8. Isolated miRNAs were marked at the 3 'end with Cy3 using the mirVana miRNA Array Labeling Kit. The miRNA profiling was done in duplicate, using microarrays containing probes for 467 mature human miRNAs.

After hybridization, the miRNA arrays were scanned using a GenePix 4000A array scanner and the raw data were normalized and analyzed using GeneSpring 7.0

Software (Silicon Genetics, Redwood City, CA). Normalization was performed expressing each miRNA replicate relative to the control microRNA (Ambion) added to each sample, allowing comparisons between the chips.

Comparisons between tumor exosomes derived from peripheral circulation and tumors indicated that miRNA signatures were not significantly different (Figure

9). This approach confirmed that at least the 12 specific miRNAs were elevated in the NSCLC and that the associations of these 12 mirrored the circulating tumor-derived exosomes.

Thus, the assessment of these miRNAs can be used as a substitute for their levels in the tumor and, therefore, are diagnoses for the presence of cancer, and in this particular case, the NSCLC.

EXAMPLE 7

PROFILE OF EXOSOMAL MYRNA DERIVED FROM

PLACENTA FOR CORRELATION WITH ADVERSE RESULTS OF PREGNANCES

To determine whether circulating exosomes comprise miRNA that can be diagnostic for adverse pregnancy outcomes (eg premature birth), serum samples were collected from pregnant individuals and exosomal fractions derived from placental tissues were isolated using antiplacental alkaline phosphatase antibodies linked with magnetic beads . The miRNAs were isolated and profiled from the isolated circulating exosomes derived from the placenta and directly from the same individual's placental tissue as revealed above in the present and shown in Figure 8. Briefly, the isolated miRNAs were labeled with Cy3 at the 3 'end using the labeling kit mirVana miRNA Array. The miRNA profiling was done in 5 duplicates, using microarrays containing probes for 467 mature human miRNA. After hybridization, the miRNA arrays were scanned using a GenePix 4000A array scanner and the raw data were normalized and analyzed using the

GeneSpring 7.0 Software (Silicon Genetics, Redwood City, CA). Normalization was done expressing each miRNA replicate relative to the control microRNA (Ambion) added to each sample, allowing comparisons between the chips.

The results are shown in Table 3. The DT1 samples are miRNA isolated from the placental tissue of women 15 who carried the pregnancy to term. DT2 samples are miRNA isolated from exosomes derived from the placenta of women who have carried the pregnancy to term. DT3 samples are miRNA isolated from the placental tissue of women who had a preterm delivery (delivery before 32 weeks of gestation). DT4 samples are miRNA isolated from exosomes derived from the placenta of women who have given birth to preterm infants. The darkened cells indicate the presence of the miRNA sample tested ...

These data demonstrate that the miRNA profiling of the exosomes derived from the placenta was obtained and that these 25 data correlate with the miRNA profiles of the placenta. Thus, miRNA profiles of miRNA isolated from exosomes produced by placental cells can be used for diagnostic purposes of adverse pregnancy outcomes.

TABLE 3

DETECTION AND QUANTIFICATION OF EXOSOME miRNA

PERIPHERAL PLACENTARIES DERIVED FROM BLOOD AND PLACENTARY TISSUE

RELATED

<td>Normalized Threshold _:</td><td></td><td> 3,11</td><td> 3,14</td><td> 4,26</td><td> 4,85</td>

<td>Standardized TPT95 /</td><td></td><td> 4,61</td><td> 5,07</td><td> 5,53</td><td> 5,20</td>

<td></td><td></td><td>DT1</td><td>DT2</td><td>DT3</td><td>DT4</td>

<td>Name</td><td>ID</td><td>8 66A</td><td>866B</td><td>866C</td><td>866D</td>

<td>hsa-miR-296</td><td> 1098</td><td> 1 5,05</td><td> 7 4,33</td><td> 4,24</td><td> 4,79</td>

<td>hsa-miR-330</td><td> 1002</td><td> 2,98</td><td> 3,09</td><td> 4,1</td><td> 1,08</td>

<td>hsa-miR-20a</td><td> 1007</td><td> 11,4 6</td><td> 11,35</td><td> 7..:7:-:-/ ///1:2:</td><td> : 11,93</td>

<td>hsa-miR-28</td><td> 1024</td><td></td><td> 10,05</td><td> 9,19</td><td> 9,23</td>

<td>hsa-miR-302c</td><td> 1032</td><td> -0,58</td><td> 3,08</td><td> 3,5</td><td> 1,08</td>

<td>hsa-miR-302a</td><td> 1036</td><td> 2,17</td><td> 7 3,66</td><td> 3,47</td><td> 4,33</td>

<td>hsa-miR-214</td><td> 1057</td><td> 6, 58</td><td> / 3, 93</td><td> 6,17</td><td> / 2,99</td>

<td>hsa-miR-99b</td><td> 1063</td><td> 9, 59</td><td> 10,08</td><td> 7 / 9, 8:6/</td><td> 9 , 16</td>

<td>hsa-miR-99a</td><td> 1068</td><td> 3,81</td><td> 7 4:,53</td><td> -7//34:</td><td> 7 7/6, 46</td>

<td>hsa-miR-lOa</td><td> 1072</td><td> 10,1</td><td> 10,76</td><td> 9, 63</td><td> 9,55</td>

<td>hsa-let-7d</td><td> 1085</td><td> 12,53</td><td> 13,32</td><td> 7 12,65</td><td> 12,5</td>

<td>hsa-miR-138</td><td> 1089</td><td> &gt; 5,37</td><td> 7 5,26</td><td> 4,18</td><td> 3,61</td>

<td>hsa-miR-140</td><td> 1106</td><td> 3,23</td><td> / 4,3</td><td> 2,01</td><td> 0,58</td>

<td>hsa-miR-23a</td><td> 1114</td><td> 14,51</td><td> 15,08</td><td> 14,99</td><td> 14,78</td>

<td>hsa-miR-215</td><td> 1122</td><td> 0,71</td><td> 1,79</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-183</td><td> 1127</td><td> 9,08</td><td> 9, 63</td><td> /:: 8,99</td><td> 8, 9</td>

<td>hsa-miR-32</td><td> 1135</td><td> 2</td><td> 2,49</td><td> 2,42</td><td> 0,58</td>

<td>hsa-miR-25</td><td> 1139</td><td> 7 11,34</td><td> 7 7 11,:.3</td><td> :. 12,23</td><td> 12,01</td>

<td>hsa-miR-218</td><td> 1143</td><td> 2,71</td><td> ::3,37</td><td> ,. 4,61</td><td> 5,33</td>

<td>hsa-miR-107</td><td> 1163</td><td> /7 11-, 68</td><td> 7 12,18</td><td> /11,29</td><td> 11,31</td>

<td>hsa-miR-145</td><td> 1168</td><td> 1,74</td><td> 2,38</td><td> 3,47</td><td> 1,38</td>

<td>hsa-miR-181a</td><td> 1172</td><td> .7/11,9</td><td> 12,62</td><td> 11,35</td><td>: vii, is</td>

<td>hsa-miR-125a</td><td> 1193</td><td> 12,34</td><td> 13,07</td><td> 11, 67</td><td> 7,11,84</td>

<td>hsa-miR-222</td><td> 1198</td><td> 12,37</td><td> 1:2:,53</td><td> / 11,77</td><td> 10,99</td>

<td>hsa-miR-372</td><td> 1105</td><td> -0,58</td><td> 3,08</td><td> 2,51</td><td> 1,08</td>

<td>hsa-miR-9 *</td><td> 1232</td><td> -0,58</td><td> -0,01</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-142-3p</td><td> 1253</td><td> 2,72</td><td> 3,06</td><td> 4,59</td><td> 3,91</td>

<td>hsa-miR-198</td><td> 1258</td><td> 7. 4,2</td><td> 7 3 , .92</td><td> 3,32</td><td> 3,67</td>

<td>hsa-miR-196a</td><td> 1263</td><td> 7 : 4., 78:</td><td> 7-/5/, 07</td><td> 3,42</td><td> 4,04</td>

<td>hsa-miR-16</td><td> 1272</td><td> 12,28</td><td> 12,05</td><td> 12,98</td><td> 12, 6</td>

<td>hsa-miR-200a</td><td> 1287</td><td> 8,29</td><td> 8,72</td><td> 7/7:,::17-</td><td> 7,44</td>

<td>hsa-miR-18a</td><td> 1292</td><td> 7 6,41</td><td> 7 6,66</td><td> 7,98</td><td> 8,5</td>

<td>hsa-miR-101</td><td> 1297</td><td>i 4.62</td><td> 4,87</td><td> 5,55</td><td> 6,01</td>

<td>hsa-miR-195</td><td> 1311</td><td> 7: 6,09</td><td> 6,58</td><td> /. . 6,03</td><td> 7 6,43</td>

<td>hsa-miR-136</td><td> 1203</td><td> 3,69</td><td> 3,77</td><td> 3,52</td><td> 3,65</td>

<td>hsa-miR-153</td><td> 1225</td><td> 1,71</td><td> 2,08</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-31</td><td> 1227</td><td> 8, 97</td><td> 9,49</td><td> 7: 9,/6/</td><td> / 9,32</td>

<td>hsa-miR-184</td><td> 1242</td><td> -0,08</td><td> -0,51</td><td> 3,1</td><td> 0,58</td>

<td>hsa-miR-33</td><td> 1274</td><td> 2,01</td><td> 3, 66</td><td> 3,58</td><td> 3,75</td>

<td>hsa-miR-147</td><td> 1278</td><td> /4,.-65/</td><td> 2,95</td><td> 3,47</td><td> 3,17</td>

<td>hsa-miR-106b</td><td> 1282</td><td> 9,47</td><td> 7 9,19</td><td> 10,59</td><td> - 10,38</td>

<td>hsa-miR-212</td><td> 1288</td><td> 2,67</td><td> 1,81</td><td> 2,89</td><td> 3,75</td>

<td>hsa-miR-15a</td><td> 1312</td><td> 9,92</td><td> 10,12</td><td> :10,79</td><td> 11,06</td>

<td>hsa-miR-135b</td><td> 1331</td><td> . 4,51</td><td> 4,03</td><td> 4,42</td><td> 4,07</td>

<td>hsa-miR-92</td><td> 1335</td><td> 12,29</td><td> 12, 63</td><td> 12,62</td><td> 12,26</td>

<td>hsa-miR-342</td><td> 1349</td><td> . 9,14</td><td> 9, 62</td><td> 8,36</td><td> 8,3 9</td>

<td>hsa-miR-205</td><td> 1368</td><td> 6,15</td><td> 5,74</td><td> 15,25</td><td> 14,33</td>

<td>hsa-miR-150</td><td> 1385</td><td> 4,02</td><td> 2,87</td><td> 3,33</td><td> 1,38</td>

<td>hsa-miR-149</td><td> 1392</td><td> 6,23</td><td> 5,65</td><td> 5, 92</td><td> 4,79</td>

<td>hsa-let-7b</td><td> 1431</td><td> 12,73</td><td> 12,43</td><td> 13,73</td><td> 14,06</td>

<td>hsa-miR-370</td><td> 1445</td><td> 4,14</td><td> 3,7</td><td> / 4,37</td><td> 2,58</td>

<td>hsa-miR-206</td><td> 1449</td><td> 5,22</td><td> 4,34</td><td> 5,5.8</td><td> 4,91</td>

<td>hsa-miR-128a</td><td> 1350</td><td> 7,54</td><td> 7,9</td><td> 8,06</td><td> 8,01</td>

<td>hsa-miR-197</td><td> 1487</td><td> 11,05</td><td> 9,76</td><td> 10,03</td><td> 9,28</td>

<td>hsa-miR-182</td><td> 1506</td><td> 10,11</td><td> 10,89</td><td> 10,09</td><td> 10,41</td>

<td>hsa-miR-553</td><td> 1750</td><td> 2,77</td><td> 2,95</td><td> 4,05</td><td> 3,17</td>

<td>hsa-miR-606</td><td> 1758</td><td> -0,58</td><td> -0,51</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-518f</td><td> 1762</td><td> 1</td><td> -0,01</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-519d</td><td> 1766</td><td> 2,17</td><td> 3,28</td><td> 0,51</td><td> 3,04</td>

<td>hsa-miR-520g</td><td> 1770</td><td> -0,58</td><td> 1,9</td><td> 3,51</td><td> 2,49</td>

<td>hsa-miR-520e</td><td> 1774</td><td> -0,58</td><td> 1,79</td><td> 0,51</td><td> 3,75</td>

<td>hsa-miR-329</td><td> 1778</td><td> 2,67</td><td> 2,08</td><td> 3,32</td><td> 2,91</td>

<td>hsa-miR-634</td><td> 1782</td><td> 6,69</td><td> 2,64</td><td> 3,32</td><td> 1,08</td>

<td>hsa-miR-614</td><td> 1786</td><td> 1,21</td><td> -0,01</td><td> 1,31</td><td> 2,91</td>

<td>hsa-miR-596</td><td> 1794</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-182 *</td><td> 1457</td><td> -0,08</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-223</td><td> 1467</td><td> 2,71</td><td> 2,49</td><td> 4,17</td><td> 3,17</td>

<td>hsa-miR-512-5p</td><td> 1843</td><td> -0,58</td><td> 0,79</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-643</td><td> 1855</td><td> -0,08</td><td> -0,51</td><td> 0,51</td><td> 2,91</td>

<td>hsa-miR-591</td><td> 1859</td><td> 0,21</td><td> 1,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-485-5p</td><td> 1863</td><td> 4,27</td><td> 2,49</td><td> 2,6</td><td> 0,58</td>

<td>hsa-miR-369-5p</td><td> 1867</td><td> 2</td><td> 1,49</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-575</td><td> 1871</td><td> 2,75</td><td> 2,69</td><td> 4,36</td><td> 3,75</td>

<td>hsa-miR-626</td><td> 1879</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-650</td><td> 1883</td><td> -0,08</td><td> -0,51</td><td> 3,68</td><td> 0,58</td>

<td>hsa-miR-663</td><td> 1891</td><td> 5,41</td><td> :////^;P5////:</td><td> 6,17</td><td> /:. 5,7 6</td>

<td>hsa-miR-520f hsamiR-520c</td><td> 1802</td><td> 1, 61</td><td> 1,79</td><td> 2,97</td><td> 3,15</td>

<td>hsa-miR-382</td><td> 1806</td><td> 4,48</td><td> 4,14</td><td> 4,04</td><td> 3,25</td>

<td>hsa-miR-656</td><td> 1810</td><td> -0,58</td><td> 0,29</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-605</td><td> 1814</td><td> 1,08</td><td> -0,51</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-655</td><td> 1920</td><td> 1,21</td><td> 2,3</td><td> 2,1</td><td> 3,17</td>

<td>hsa-miR-545</td><td> 1932</td><td> 2,5</td><td> 2,66</td><td> 2,92</td><td> 3,58</td>

<td>hsa-miR-502</td><td> 1940</td><td> . 3,4 6</td><td> 4,16</td><td> 4,99</td><td> 3,75</td>

<td>hsa-miR-200a *</td><td> 1952</td><td> 5,35</td><td> 5,86</td><td> 3, 42</td><td> 2,25</td>

<td>hsa-miR-640</td><td> 1956</td><td> 2,24</td><td> -0,51</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-620</td><td> 1960</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 1,08</td>

<td>hsa-miR-514</td><td> 1972</td><td> 2</td><td> 2,95</td><td> 1,01</td><td> 1,38</td>

<td>hsa-miR-583</td><td> 1980</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-548b</td><td> 1988</td><td> 1,92</td><td> -0,01</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-609</td><td> 1899</td><td> 2,55</td><td> 2,58</td><td> 3, 6</td><td> 3,54</td>

<td>hsa-miR-563</td><td> 1903</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-377</td><td> 1929</td><td> 1,74</td><td> -0,01</td><td> 2,6</td><td> 0,58</td>

<td>hsa-miR-37 6a</td><td> 1933</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-433</td><td> 1937</td><td> 2,71</td><td> 2,19</td><td> 3,74</td><td> 1,08</td>

<td>hsa-miR-500</td><td> 1957</td><td> 4,67</td><td> 4,88</td><td> . 6,34</td><td> / 5,8</td>

<td>hsa-miR-652</td><td> 1961</td><td></td><td> .6,6</td><td> 5, 52</td><td> 5,05</td>

<td>hsa-miR-384</td><td> 1969</td><td> -0,58</td><td> 2,3</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-548d</td><td> 1977</td><td> -0,58</td><td> 2,3</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-518c</td><td> 1981</td><td> 0, 92</td><td> 1,81</td><td> 3, 68</td><td> 1,38</td>

<td>hsa-miR-561</td><td> 1985</td><td> -0,58</td><td> 2,49</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-551a</td><td> 2018</td><td> 3,77</td><td> 3,27</td><td> 4,06</td><td> 3,91</td>

<td>hsa-miR-554</td><td> 2026</td><td> -0,08</td><td> 0,29</td><td> 1,01</td><td> 1,08</td>

<td>hsa-miR-510</td><td> 2030</td><td> 2,59</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-525</td><td> 2034</td><td> -0,58</td><td> 2,06</td><td> 3,1</td><td> 0,58</td>

<td>hsa-miR-570</td><td> 2054</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-617</td><td> 2062</td><td> 2</td><td> 2,49</td><td> 0, 51</td><td> 2,91</td>

<td>hsa-miR-556</td><td> 2070</td><td> -0,58</td><td> 1,81</td><td> 1,31</td><td> 1,08</td>

<td>hsa-miR-551b</td><td> 2074</td><td> 1,37</td><td> 1,79</td><td> 3, 97</td><td> 3,38</td>

<td>hsa-miR-424</td><td> 1993</td><td> 5,87</td><td> 5,34</td><td> 4,54</td><td> 4,79</td>

<td>hsa-miR-612</td><td> 1997</td><td> 2,87</td><td> 0,29</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-130a</td><td> 2005</td><td> 8,07</td><td> 7 8,66</td><td> 79,087</td><td>^Ί 9,2</td>

<td>hsa-miR-569</td><td> 2110</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-302a *</td><td> 2114</td><td> -0,58</td><td> 1,95</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-499</td><td> 2122</td><td> -0,58</td><td> -0,01</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-429</td><td> 2134</td><td>V 5.68</td><td> 5,39</td><td> 3,89</td><td> 4,83</td>

<td>hsa-miR-365</td><td> 2138</td><td> 8,7</td><td> 7 7: 8,7 3</td><td> 7,51</td><td> 7,31</td>

<td>hsa-miR-598</td><td> 2150</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-29a</td><td> 2154</td><td> 13, 45</td><td> 13,83</td><td> 12,21</td><td> 12,27</td>

<td>hsa-miR-503</td><td> 2162</td><td> : ...&lt; 5,4 4</td><td> 6,25</td><td> 1,31</td><td> 4,39</td>

<td>hsa-miR-624</td><td> 2166</td><td> -0,58</td><td> 1,99</td><td> 0,51</td><td> 3,39</td>

<td>hsa-miR-409-5p</td><td> 2089</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-550</td><td> 2097</td><td> 7 8 4,34</td><td> 7 4,2 6</td><td> 3, 89</td><td> 2,58</td>

<td>hsa-miR-627</td><td> 2101</td><td> -0,58</td><td> 1,49</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-33b</td><td> 2105</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-581</td><td> 2227</td><td> 2,32</td><td> 1, 65</td><td> 2,31</td><td> 0,58</td>

<td>hsa-miR-493-3p</td><td> 2231</td><td> 2,17</td><td> 0,29</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-610</td><td> 2239</td><td> '77774:,757:</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-516-3p</td><td> 2259</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-422a</td><td> 2263</td><td> 7 7 78:,:33</td><td> 8,48</td><td> 9,59</td><td> /7 9,25</td>

<td>hsa-miR-449</td><td> 2267</td><td> 2,91</td><td> 2,48</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-585</td><td> 2271</td><td> 3,58</td><td> 3,74</td><td> 4, 51</td><td> 4,12</td>

<td>hsa-miR-379</td><td> 2275</td><td> 2,22</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-92b</td><td> 2182</td><td> 77 7,86</td><td> 778,04</td><td> 7,75</td><td> -/ 7,:13-</td>

<td>hsa-miR-629</td><td> 2316</td><td> ¢,12</td><td> 5,93</td><td> 6,82</td><td> 7 7,03</td>

<td>hsa-miR-580</td><td> 2320</td><td> -0,58</td><td> 1,49</td><td> 0, 51</td><td> 2,58</td>

<td>hsa-miR-448</td><td> 2324</td><td> 1,74</td><td> -0,51</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-659</td><td> 2328</td><td> -0,58</td><td> 0,29</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-592</td><td> 2332</td><td> 0,21</td><td> 2,95</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-587</td><td> 2336</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-671</td><td> 2839</td><td> 4,15</td><td> 3,98</td><td> 4,21</td><td> 0,58</td>

<td>hsa-miR-802</td><td> 2851</td><td> 1,42</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-767-3p</td><td> 2863</td><td> 1,42</td><td> 2,3</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-608</td><td> 2279</td><td> 78 3,74</td><td> 1,29</td><td> 2,47</td><td> 0,58</td>

<td>hsa-miR-668</td><td> 2287</td><td> 1,21</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-210</td><td> 2291</td><td> 9,13</td><td> 8,6</td><td> / 8,39</td><td> 7 7,96</td>

<td>hsa-miR-26a</td><td> 2299</td><td> 12,6</td><td> 12,61</td><td> / 12,27</td><td> 12,7 3</td>

<td>hsa-miR-493-5p</td><td> 2329</td><td> 2</td><td> 2,65</td><td> 2,17</td><td> 1,08</td>

<td>hsa-miR-202 *</td><td> 2337</td><td> 2,55</td><td> 2,08</td><td> 3, 32</td><td> 1,88</td>

<td>hsa-miR-454-5p</td><td> 2840</td><td> 7 11,:51</td><td> : 11/6</td><td> 12,87</td><td> 13,07</td>

<td>hsa-miR-770-5p</td><td> 2844</td><td> 2,24</td><td> -0,01</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-767-5p</td><td> 2848</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-769-3p</td><td> 2865</td><td> 3,8</td><td> 3,74</td><td> 3, 01</td><td> 2,88</td>

<td>hsa-miR-758</td><td> 2869</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-765</td><td> 2864</td><td> 5,35</td><td> 5,13</td><td> 6,08</td><td> ,. 5,77</td>

<td>hsa-miR-301</td><td> 1103</td><td> 3,96</td><td> 4,08</td><td> 4,04</td><td> 4,68</td>

<td>hsa-miR-191</td><td> 1017</td><td> 9,84</td><td> 10,87</td><td> 11,01</td><td> 10,9</td>

<td>hsa-miR-93</td><td> 1029</td><td> 9, 68</td><td> 9,58</td><td> 10,93</td><td> 9,75</td>

<td>hsa-let-7f</td><td> 1033</td><td> 11,59</td><td> 12,43</td><td> 11,79</td><td> 11,9 4</td>

<td>hsa-miR-373</td><td> 1037</td><td> 3,54</td><td> 2,4</td><td> 4,51</td><td> 3,08</td>

<td>hsa-miR-200b</td><td> 1042</td><td> 10,9</td><td>/ n, 07</td><td> 10,17</td><td> 9,02</td>

<td>hsa-miR-100</td><td> 1064</td><td> 7,25</td><td> 6, 69</td><td> . 7,81</td><td> 5,54</td>

<td>hsa-miR-324-3p</td><td> 1082</td><td> 5,67</td><td> 4,71</td><td> 5, 64</td><td> 3,99</td>

<td>hsa-miR-34b</td><td> 1096</td><td> 3,27</td><td> :/· 3,49</td><td> 3,83</td><td> 4,54</td>

<td>hsa-miR-324-5p</td><td> 1115</td><td> 3,84</td><td> 2,29</td><td> 4,16</td><td> 4,49</td>

<td>hsa-miR-199a *</td><td> 1124</td><td> 1,82</td><td> 2,24</td><td> 1,01</td><td> 4,17</td>

<td>hsa-miR-103</td><td> 1164</td><td> 11,27</td><td> 10, 65</td><td> 11,3</td><td> 7^:; 9,18</td>

<td>hsa-miR-142-5p</td><td> 1169</td><td> -0,58</td><td> -0,51</td><td> 2,31</td><td> 0,58</td>

<td>hsa-miR-220</td><td> 1173</td><td> 3,67</td><td> 3,04</td><td> 4,26</td><td> 3,99</td>

<td>hsa-miR-151</td><td> 1199</td><td> ···. 9,73</td><td> / 9,47</td><td> 10,22</td><td> 10,45</td>

<td>hsa-miR-18 6</td><td> 1141</td><td> /. 4,72</td><td> 4,93</td><td> 3,86</td><td> 4,49</td>

<td>hsa-miR-128b</td><td> 1153</td><td> 6,29</td><td> /76,26</td><td> -77/:/6/77</td><td> 77: 6,1.</td>

<td>hsa-miR-130b</td><td> 1165</td><td> 7,72</td><td> 7 6, 96</td><td> 7,99</td><td> / 6,49</td>

<td>hsa-miR-338</td><td> 1174</td><td> 2,42</td><td> 2,66</td><td> 2,67</td><td> 2,91</td>

<td>hsa-miR-199b</td><td> 1178</td><td> 1,98</td><td> -0,01</td><td> 3, 67</td><td> 3,46</td>

<td>hsa-miR-125b</td><td> 1182</td><td> 9,34</td><td> 8,81</td><td> 9,8 6</td><td> 8,11</td>

<td>hsa-miR-124a</td><td> 1213</td><td> 1,74</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-122a</td><td> 1243</td><td> /; 5,11</td><td> 3,49</td><td> ? 4,97.</td><td> 4,71</td>

<td>hsa-miR-30d</td><td> 1251</td><td> 11,72</td><td> 11, 93</td><td> 11,32</td><td> 11,69</td>

<td>hsa-miR-203</td><td> 1260</td><td> 1,42</td><td> 2,9</td><td> 9,1</td><td> 9,56</td>

<td>hsa-let-7c</td><td> 1268</td><td> 11, 91</td><td> 12,72</td><td> /:13,09</td><td> 12,47</td>

<td>hsa-miR-216</td><td> 1294</td><td> 2</td><td> 2,45</td><td> 2,71</td><td> 3,38</td>

<td>hsa-miR-144</td><td> 1300</td><td> 0,71</td><td> 0,49</td><td> 1,01</td><td> 2,91</td>

<td>hsa-miR-15b</td><td> 1313</td><td> 11,75</td><td> 12,27</td><td> 12,66</td><td> 12,77</td>

<td>hsa-miR-192</td><td> 1205</td><td> 7,05</td><td> 8,48</td><td> .//,/6://</td><td> . 6,14</td>

<td>hsa-miR-133a</td><td> 1215</td><td> , 3,2 7</td><td> 3,07</td><td> 3,82</td><td> 4,11</td>

<td>hsa-miR-126</td><td> 1380</td><td> ./. 6, 42</td><td> 6,42</td><td> 5,94</td><td> 77,51</td>

<td>hsa-miR-326</td><td> 1393</td><td> 3, 32</td><td> 0,29</td><td> 0,51</td><td> 3,17</td>

<td>hsa-miR-98</td><td> 1423</td><td> 6,58</td><td> 7,21</td><td> 6,9</td><td> 7/ 7,33</td>

<td>hsa-let-7g</td><td> 1432</td><td> 10,8</td><td> 7 11,21</td><td> 7. 10,01</td><td> / 10,06</td>

<td>hsa-miR-190</td><td> 1437</td><td> 3,16</td><td> 3,57</td><td> 4,02</td><td> 4,29</td>

<td>hsa-miR-189</td><td> 1442</td><td> 2,59</td><td> 2,79</td><td> 2,92</td><td> 3,38</td>

<td>hsa-miR-137</td><td> 1339</td><td> 2,66</td><td> 3,06</td><td> / 4,36</td><td> 3,88</td>

<td>hsa-miR-105</td><td> 1345</td><td> 2,37</td><td> 2,48</td><td> 74,32</td><td> 3,17</td>

<td>hsa-miR-96</td><td> 1507</td><td> 4,66</td><td> 4,1.7</td><td> 4,58</td><td> 4,58</td>

<td>hsa-miR-518b</td><td> 1759</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-519e</td><td> 1767</td><td> -0,08</td><td> 2,3</td><td> 2,83</td><td> 1,08</td>

<td>hsa-miR-520a</td><td> 1771</td><td> 1,42</td><td> 1,99</td><td> 3,1</td><td> 0,58</td>

<td>hsa-miR-552</td><td> 1779</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-451</td><td> 1783</td><td> 1</td><td> -0,51</td><td> 3,32</td><td> 2,58</td>

<td>hsa-miR-523</td><td> 1787</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-518e</td><td> 1795</td><td> -0,08</td><td> -0,51</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-299-5p</td><td> 1458</td><td> 1,74</td><td> 2,06</td><td> 2,51</td><td> 0,58</td>

<td>hsa-miR-95</td><td> 1482</td><td> 3,94</td><td> 3,78</td><td> 2,71</td><td> 3,08</td>

<td>hsa-miR-520h</td><td> 1824</td><td> 1,74</td><td> 1,08</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-593</td><td> 1832</td><td> 7 4,92</td><td> 1,08</td><td> 4,1</td><td> 2,91</td>

<td>hsa-miR-57 4</td><td> 1840</td><td> 11,34</td><td> 9,36</td><td> 11,12</td><td> 9, 45</td>

<td>hsa-miR-641</td><td> 1856</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-504</td><td> 1860</td><td> -0,58</td><td> 1,81</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-202</td><td> 1864</td><td> 2,59</td><td> 1,9</td><td> 2,97</td><td> 3, 67</td>

<td>hsa-miR-564</td><td> 1884</td><td> 0, 42</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-604</td><td> 1892</td><td> 1,71</td><td> -0,51</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-519b</td><td> 1799</td><td> -0,08</td><td> 3,49</td><td> 3,97</td><td> 3,83</td>

<td>hsa-miR-520d</td><td> 1803</td><td> 2,58</td><td> 3, 56</td><td> 3,89</td><td> 4,36</td>

<td>hsa-miR-602</td><td> 1825</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-622</td><td> 1829</td><td> 1,42</td><td> 2,66</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-483</td><td> 1845</td><td> 8,06</td><td> 3,91</td><td> 7 7,36</td><td> 4,34</td>

<td>hsa-miR-600</td><td> 1853</td><td> 0,21</td><td> -0,01</td><td> 0,51</td><td> 2,38</td>

<td>hsa-miR-631</td><td> 1861</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-383</td><td> 1865</td><td> 2,17</td><td> 1,95</td><td> 0,51</td><td> 1,88</td>

<td>hsa-miR-29b</td><td> 1869</td><td> 6, 67</td><td> 6,05</td><td> 5,9</td><td> 7: 6, 65</td>

<td>hsa-miR-613</td><td> 1881</td><td> 2,42</td><td> -0,01</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-453</td><td> 1904</td><td> 7 3,93</td><td> 3,59</td><td> 4,76</td><td> /7/4., 17</td>

<td>hsa-miR-489</td><td> 1908</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-23b</td><td> 1930</td><td> •;3,2</td><td> 13,31</td><td> 7 12,99</td><td> 13, 51</td>

<td>hsa-miR-376b</td><td> 1934</td><td> -0,58</td><td> -0,01</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-501</td><td> 1942</td><td> 2,87</td><td> 2,72</td><td> 1,31</td><td> 3,25</td>

<td>hsa-miR-517c</td><td> 1946</td><td> 3,01</td><td> 3,02</td><td> 4,36</td><td> 4,39</td>

<td>hsa-miR-516-5p</td><td> 1950</td><td> -0,08</td><td> -0,51</td><td> 1, 01</td><td> 0,58</td>

<td>hsa-miR-548c</td><td> 1978</td><td> -0,58</td><td> -0,51</td><td> 1,01</td><td> 1,08</td>

<td>hsa-miR-625</td><td> 1897</td><td> 6,54</td><td> 7,16</td><td> 5, 67</td><td> 5,55</td>

<td>hsa-miR-630</td><td> 1905</td><td> 2,74</td><td> 1,49</td><td> 2, 92</td><td> 3,49</td>

<td>hsa-miR-644</td><td> 1913</td><td> 0,21</td><td> 2,29</td><td> 2,51</td><td> 2,91</td>

<td>hsa-miR-488</td><td> 2015</td><td> -0,58</td><td> 0,29</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-633</td><td> 2023</td><td> 1,42</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-527</td><td> 2039</td><td> .··: 3,42</td><td> 1,87</td><td> 2,51</td><td> 1,08</td>

<td>hsa-miR-589</td><td> 2055</td><td> -0,08</td><td> -0,51</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-508</td><td> 2071</td><td> 2,81</td><td> 2,79</td><td> 3,21</td><td> 4,04</td>

<td>hsa-miR-566</td><td> 2075</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-449b</td><td> 2083</td><td> 2,41</td><td> 2,48</td><td> 3, 97</td><td> 2,58</td>

<td>hsa-miR-603</td><td> 1990</td><td> 7 4,74</td><td> 2,84</td><td> 3,76</td><td> 0,58</td>

<td>hsa-miR-607</td><td> 2111</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-559</td><td> 2115</td><td> -0,58</td><td> 0,29</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-506</td><td> 2123</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-200c</td><td> 2131</td><td> 4,25</td><td> 3,7 5</td><td> 7 13,3</td><td> 13,7</td>

<td>hsa-miR-29c</td><td> 2155</td><td> 2,75</td><td> 3,24</td><td> 3, 6</td><td> 4,2</td>

<td>hsa-miR-411</td><td> 2167</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-381</td><td> 2171</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-455</td><td> 2179</td><td> 2,87</td><td> 2,52</td><td> 2,6</td><td> 0,58</td>

<td>hsa-miR-363 *</td><td> 2086</td><td> 1,42</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-380-5p</td><td> 2090</td><td> 0,21</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-567</td><td> 2094</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-378</td><td> 2208</td><td> 2,24</td><td> 2,3</td><td> 3, 6</td><td> 2,58</td>

<td>hsa-miR-638</td><td> 2212</td><td> 7,37</td><td> 6, 34</td><td> 7 7 8,56</td><td> 77' 7,57</td>

<td>hsa-miR-542-5p</td><td> 2216</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-518f *</td><td> 2220</td><td> -0,58</td><td> -0,51</td><td> 3, 17</td><td> 3, 67</td>

<td>hsa-miR-54 9</td><td> 2232</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-558</td><td> 2240</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-let-7i</td><td> 2244</td><td> 12,8</td><td> 13,03</td><td> 10,86</td><td> 710,79</td>

<td>hsa-miR-560</td><td> 2256</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-636</td><td> 2260</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 3,17</td>

<td>hsa-miR-422b</td><td> 2264</td><td> 9,17</td><td> 7 8,72</td><td> 7 10,51</td><td> 7 10,23</td>

<td>hsa-miR-193b</td><td> 2268</td><td> 9, 68</td><td> 8,44</td><td> 8, 63</td><td> 7,54</td>

<td>hsa-miR-491</td><td> 2272</td><td> 1,74</td><td> 0,79</td><td> 2,81</td><td> 0,58</td>

<td>hsa-miR-484</td><td> 2191</td><td> 8,32</td><td> 7,81</td><td> 8,29</td><td> 7,72</td>

<td>hsa-miR-662</td><td> 2199</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-623</td><td> 2203</td><td> 1,74</td><td> 2,45</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-486</td><td> 2209</td><td> 3,86</td><td> 3,3</td><td> 4,2</td><td> 4,6</td>

<td>hsa-miR-639</td><td> 2213</td><td> 1,87</td><td> 1,49</td><td> 2,31</td><td> 1,38</td>

<td>hsa-miR-517a hsamiR-517b</td><td> 2217</td><td> 2,11</td><td> 2,56</td><td> 3,87</td><td> 3,28</td>

<td>hsa-miR-645</td><td> 2221</td><td> 3, 12</td><td> 1,29</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-653</td><td> 2229</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-146b</td><td> 2237</td><td> 5,56</td><td> 5,2.9</td><td> 4,21</td><td> 5,59</td>

<td>hsa-miR-571</td><td> 2249</td><td> 3,33</td><td> 2,99</td><td> 4,1</td><td> 2,91</td>

<td>hsa-miR-191 *</td><td> 2257</td><td> 2,42</td><td> 2, 95</td><td> 1,31</td><td> 0, 58</td>

<td>hsa-miR-7</td><td> 2261</td><td> 2,44</td><td> 3,02</td><td> 2,51</td><td> 3,54</td>

<td>hsa-miR-647</td><td> 2269</td><td> 4,95</td><td> 4,27</td><td> / 5,5</td><td> 6, 01</td>

<td>hsa-miR-637</td><td> 2273</td><td> .. 4,65</td><td> 2,84</td><td> 4,9</td><td> 4,17</td>

<td>hsa-miR-30b</td><td> 2280</td><td> 9,94</td><td> 9,87</td><td> / 9,8 6</td><td> 9, 66</td>

<td>hsa-miR-431</td><td> 2288</td><td> 1,74</td><td> -0,01</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-452</td><td> 2292</td><td> 4,68</td><td> 5,15</td><td> 5,14</td><td> 5,85</td>

<td>hsa-miR-361</td><td> 2296</td><td> 10,36</td><td> 11,32</td><td> 10,53</td><td> 10,8.3</td>

<td>hsa-miR-576</td><td> 2314</td><td> 1,87</td><td> -0,51</td><td> 2,83</td><td> 0,58</td>

<td>hsa-miR-432</td><td> 2326</td><td> 3,74</td><td> 3,47</td><td> 3, 51</td><td> 2,58</td>

<td>hsa-miR-375</td><td> 2342</td><td> 3,42</td><td> 2,15</td><td> 0, 51</td><td> 3,75</td>

<td>hsa-miR-766</td><td> 2841</td><td> 9, 66</td><td> 6,37</td><td> 8,18</td><td> 7,59</td>

<td>hsa-miR-768-3p</td><td> 2845</td><td> 9,89</td><td> 9,61</td><td> 9,2</td><td> 9,48</td>

<td>hsa-miR-769-5p</td><td> 2861</td><td> 4,03</td><td> 4,07</td><td> 4,47</td><td> 3,46</td>

<td>hsa-miR-542-3p</td><td> 2289</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-513</td><td> 2301</td><td> 3,8</td><td> 2,56</td><td> 3, 97</td><td> 4,38</td>

<td>hsa-miR-362</td><td> 2017</td><td> 2,93</td><td> 4,53</td><td> 4,88</td><td> 4,38</td>

<td>hsa-miR-325</td><td> 2025</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-520a *</td><td> 2033</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-517 *</td><td> 2037</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-565</td><td> 2045</td><td> 7,04</td><td> 4,89</td><td> 5,13</td><td> 6,45</td>

<td>hsa-miR-526b</td><td> 2049</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-30e-3p</td><td> 2053</td><td> 8,97</td><td> 9,4</td><td> 7,8 4</td><td> 7,61</td>

<td>hsa-miR-601</td><td> 2088</td><td> 2,87</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-519a</td><td> 2104</td><td> -0,58</td><td> 1,49</td><td> 0, 51</td><td> 1,08</td>

<td>hsa-miR-632</td><td> 2108</td><td> -0,08</td><td> 2,3</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-320</td><td> 1005</td><td> 12,75</td><td> 13,28</td><td> 13,11</td><td> 13, 09</td>

<td>hsa-miR-132</td><td> 1014</td><td> 4,94</td><td> 6,57</td><td> 6,22</td><td> 7,04</td>

<td>hsa-miR-193a</td><td> 1018</td><td> 4,56</td><td> 4,32</td><td> 3, 66</td><td> 4,58</td>

<td>hsa-miR-22</td><td> 1022</td><td> 8,71</td><td> 8,95</td><td> 8, 69</td><td> 8,79</td>

<td>hsa-miR-224</td><td> 1026</td><td> 6, 69</td><td> 7,1</td><td> 6,4</td><td> 6,96</td>

<td>hsa-let-7a</td><td> 1030</td><td> 13,37</td><td> 14,07</td><td> 14, 63</td><td> 14,91</td>

<td>hsa-miR-302d</td><td> 1034</td><td> 2,32</td><td> 2,74</td><td> 3,76</td><td> 3,28</td>

<td>hsa-miR-369-3p</td><td> 1038</td><td> 2,72</td><td> 2,38</td><td> 4, 68</td><td> 3,83</td>

<td>hsa-miR-154 *</td><td> 1047</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-368</td><td> 1059</td><td> 1,42</td><td> 0,49</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-373 *</td><td> 1078</td><td> -0, 58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-34c</td><td> 1095</td><td> 1,42</td><td> 1,49</td><td> 0,51</td><td> 2,99</td>

<td>hsa-miR-154</td><td> 1101</td><td> 1,61</td><td> -0,51</td><td> 1,31</td><td> 2,75</td>

<td>hsa-miR-106a</td><td> 1006</td><td> 12,01</td><td> 12,48</td><td> 12,09</td><td> 12,36</td>

<td>hsa-miR-181c</td><td> 1015</td><td> &lt; 5, 67</td><td> 6,09</td><td> 4, 64</td><td> 4,27</td>

<td>hsa-miR-17-5p</td><td> 1031</td><td> 11,57</td><td> 11,85</td><td> 11,34</td><td> 11,83</td>

<td>hsa-miR-302b</td><td> 1035</td><td> -0,08</td><td> 2,66</td><td> 3,26</td><td> 4,04</td>

<td>hsa-miR-19b</td><td> 1039</td><td> 10,14</td><td> 10,07</td><td> 11,3</td><td> 11,47</td>

<td>hsa-miR-24</td><td> 1044</td><td> 12,91</td><td> 13,2</td><td> 13,13</td><td> 13,4</td>

<td>hsa-miR-367</td><td> 1052</td><td> 2,17</td><td> -0,01</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-17-3p</td><td> 1079</td><td> 4,95</td><td> 5,02</td><td> 4,83</td><td> 5,34</td>

<td>hsa-miR-221</td><td> 1088</td><td> 13, 67</td><td> 13,73</td><td> 12,88</td><td> 12,7 6</td>

<td>hsa-miR-335</td><td> 1146</td><td> -0,58</td><td> -0,51</td><td> 6, 66</td><td> 7,68</td>

<td>hsa-miR-323</td><td> 1154</td><td> -0,58</td><td> 1,81</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-199a</td><td> 1167</td><td> 2,31</td><td> -0,51</td><td> 0,51</td><td> 3, 17</td>

<td>hsa-miR-126 *</td><td> 1171</td><td> 3,12</td><td> 1,95</td><td> 3,68</td><td> 3,15</td>

<td>hsa-miR-337</td><td> 1175</td><td> 2,22</td><td> -0,51</td><td> 3,97</td><td> 2,91</td>

<td>hsa-miR-181a *</td><td> 1179</td><td> 5,67</td><td> 5,34</td><td> 5,91</td><td> 5,76</td>

<td>hsa-miR-331</td><td> 1183</td><td> 6,46</td><td> 5,25</td><td> 5,55</td><td> 4,95</td>

<td>hsa-miR-340</td><td> 1187</td><td> 2,96</td><td> 2,99</td><td> 3,86</td><td> 4,17</td>

<td>hsa-miR-208</td><td> 1108</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-188</td><td> 1116</td><td> 3,94</td><td> 3,31</td><td> 3,86</td><td> 4,39</td>

<td>hsa-miR-9</td><td> 1231</td><td> 2,96</td><td> 3,25</td><td> 4</td><td> 4,53</td>

<td>hsa-miR-34a</td><td> 1235</td><td> 6,95</td><td> 6,56</td><td> 7,17</td><td> 7,33</td>

<td>hsa-miR-30c</td><td> 1252</td><td> 13,78</td><td> 13,97</td><td> 12,46</td><td> 12,24</td>

<td>hsa-miR-19a</td><td> 1271</td><td> 5,93</td><td> 5,7 6</td><td> 8,01</td><td> ,, . 8,36</td>

<td>hsa-miR-371</td><td> 1276</td><td> 3,67</td><td> 2,19</td><td> 3,36</td><td> 3,38</td>

<td>hsa-miR-lOb</td><td> 1301</td><td> 6,91</td><td> 7,36</td><td> 7,73</td><td> 8,03</td>

<td>hsa-miR-21</td><td> 1315</td><td> 13,13</td><td> 13,2</td><td> 12,28</td><td> ,12,88</td>

<td>hsa-miR-217</td><td> 1206</td><td> 2,53</td><td> 2,49</td><td> 0,51</td><td> 3,57</td>

<td>hsa-miR-302b *</td><td> 1210</td><td> 1,87</td><td> 2,49</td><td> 2,51</td><td> 2,99</td>

<td>hsa-miR-135a</td><td> 1216</td><td> 2,41</td><td> 3, 62</td><td> 3,47</td><td> 3,89</td>

<td>hsa-miR-148a</td><td> 1361</td><td> 3</td><td> 1,45</td><td> 6,87</td><td> 7,35</td>

<td>hsa-miR-339</td><td> 1366</td><td> 4,85</td><td> 4,26</td><td> 5,12</td><td> 5,2</td>

<td>hsa-miR-187</td><td> 1381</td><td> 3,69</td><td> 2,4</td><td> 4,21</td><td> 3,75</td>

<td>hsa-miR-34 6</td><td> 1390</td><td> 5,77</td><td> 3,2</td><td> 4,09</td><td> 4,87</td>

<td>hsa-miR-146a</td><td> 1409</td><td> 9,7</td><td> 9,88</td><td> 7,17</td><td> . 7,56</td>

<td>hsa-miR-143</td><td> 1415</td><td> -0,58</td><td> -0,51</td><td> 2,51</td><td> 3,75</td>

<td>hsa-miR-219</td><td> 1426</td><td> 2</td><td> 1,81</td><td> 3,32</td><td> 4,04</td>

<td>hsa-miR-185</td><td> 1451</td><td> 00</td><td> 8,73</td><td> 9,33</td><td> 9,46</td>

<td>hsa-miR-328</td><td> 1455</td><td> 7,15</td><td> 4,5</td><td> 4,92</td><td> 4,33</td>

<td>hsa-miR-196b</td><td> 1321</td><td> ' 4,65</td><td> 4,44</td><td> 5,08</td><td> 5, 68</td>

<td>hsa-miR-204</td><td> 1489</td><td> 0,71</td><td> 2,49</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-133b</td><td> 1498</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-129</td><td> 1512</td><td> 6,33</td><td> 6,08</td><td> 7,2</td><td> 8,02</td>

<td>hsa-miR-649</td><td> 1756</td><td> 3,32</td><td> 2,93</td><td> 3,17</td><td> 2,17</td>

<td>hsa-miR-518a</td><td> 1760</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-562</td><td> 1764</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 1,08</td>

<td>hsa-miR-526b *</td><td> 1772</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-522</td><td> 1776</td><td> 2,87</td><td> 3/4</td><td> 5,74</td><td> 5, 87</td>

<td>hsa-miR-4 90</td><td> 1784</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-618</td><td> 1788</td><td> 2,22</td><td> 1,65</td><td> 0,51</td><td> 1,08</td>

<td>hsa-miR-525 *</td><td> 1796</td><td> -0,58</td><td> 1,49</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-30a-5p</td><td> 1460</td><td> 12,45</td><td> 12,55</td><td> 11,09</td><td> 11,04</td>

<td>hsa-miR-302c *</td><td> 1474</td><td> 0,42</td><td> -0,01</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-27a</td><td> 1485</td><td> 11,64</td><td> 11,67</td><td> 11,97</td><td> 12,27</td>

<td>hsa-miR-30a-3p</td><td> 1505</td><td> 12,22</td><td> 12,57</td><td> 10</td><td> 10,48</td>

<td>hsa-miR-494</td><td> 1753</td><td> / 4,47</td><td> 3,87</td><td> 6,12</td><td> 5, 48</td>

<td>hsa-miR-518d</td><td> 1761</td><td> -0,58</td><td> 2,08</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-519c</td><td> 1765</td><td> -0,08</td><td> 0,29</td><td> 0,51</td><td> 3,75</td>

<td>hsa-miR-20b</td><td> 1769</td><td> 10,41</td><td> 10,8</td><td> 10,92</td><td> 11,2 .</td>

<td>hsa-miR-520b</td><td> 1773</td><td> -0, 08</td><td> 1,49</td><td> 1,01</td><td> 2,58</td>

<td>hsa-miR-495</td><td> 1777</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 2,91</td>

<td>hsa-miR-521</td><td> 1785</td><td> 3, 42</td><td> 0,49</td><td> 3,31</td><td> 3,75</td>

<td>hsa-miR-64 6</td><td> 1793</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-648</td><td> 1804</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-410</td><td> 1808</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 1,08</td>

<td>hsa-miR-487a</td><td> 1812</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-409-3p</td><td> 1820</td><td> -0,58</td><td> -0,51</td><td> 1,01</td><td> 0,58</td>

<td>hsa-miR-363</td><td> 1822</td><td> -0,58</td><td> -0, 51</td><td> 3,32</td><td> 1,08</td>

<td>hsa-miR-181 b</td><td> 1830</td><td> 11,53</td><td> 11,96</td><td> 10,84</td><td> 11,02</td>

<td>hsa-miR-616</td><td> 1842</td><td> 2,22</td><td> -0,51</td><td> 0,51</td><td> 2,49</td>

<td>laughs 3 &lt;3— llxR— 1 8 &lt;3. *</td><td> 1850</td><td> 4,52</td><td> 2, 99</td><td> 4,97</td><td> 3, 83</td>

<td>hsa-miR-635</td><td> 1854</td><td> -0,58</td><td> -0,51</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-423</td><td> 1874</td><td> 8,9</td><td> 8,85</td><td> 9,09</td><td> 8,46</td>

<td>hsa-miR-611</td><td> 1882</td><td> -0, 58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-524</td><td> 1797</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-595</td><td> 1805</td><td> 9,11</td><td> ··:· 6,55</td><td> 8,47</td><td> 6, 49</td>

<td>hsa-miR-487b</td><td> 1817</td><td> 4,65</td><td> 4,3</td><td> 5,22</td><td> 5,53</td>

<td>hsa-miR-425-3p</td><td> 1943</td><td> 4,14</td><td> : 4,02</td><td> 3,39</td><td> 3, 96</td>

<td>hsa-miR-594</td><td> 1951</td><td> 10, 94</td><td> 10,48</td><td> 11,55</td><td> 11,22</td>

<td>hsa-miR-532</td><td> 1959</td><td> 5, 87</td><td> 5, 79</td><td> 6,62</td><td> 6,14</td>

<td>hsa-miR-568</td><td> 1963</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-496</td><td> 1967</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-544</td><td> 1971</td><td> 1,08</td><td> 2,49</td><td> 1,01</td><td> 2, 91</td>

<td>hsa-miR-509</td><td> 1975</td><td> -0,08</td><td> -0,51</td><td> 0,51</td><td> 3, 75</td>

<td>hsa-miR-548a</td><td> 1979</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-658</td><td> 1894</td><td> -0,58</td><td> -0,51</td><td> 1,01</td><td> 0, 58</td>

<td>hsa-miR-555</td><td> 1898</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-657</td><td> 1902</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-512-3p</td><td> 1910</td><td> 2,56</td><td> 2,74</td><td> 4,41</td><td> 3,83</td>

<td>hsa-miR-524 *</td><td> 2024</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-515-5p</td><td> 2032</td><td> -0, 58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-526a</td><td> 2036</td><td> -0,58</td><td> -0,51</td><td> 5,78</td><td> 5, 87</td>

<td>hsa-miR-619</td><td> 2044</td><td> 2,01</td><td> 1,49</td><td> 1,01</td><td> 4,08</td>

<td>hsa-miR-578</td><td> 2048</td><td> 3,54</td><td> 2,79</td><td> 3,17</td><td> 2,38</td>

<td>hsa-miR-573</td><td> 2056</td><td> -0,58</td><td> 2,08</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-492</td><td> 2060</td><td> -0,08</td><td> 1,49</td><td> 2,71</td><td> 2, 67</td>

<td>hsa-miR-590</td><td> 2064</td><td> 3,27</td><td> 3,4</td><td> 5,51</td><td> 5, 08</td>

<td>hsa-miR-515-3p</td><td> 2068</td><td> 1,74</td><td> 2,88</td><td> 3, 51</td><td> 1,08</td>

<td>hsa-miR-621</td><td> 2076</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-539</td><td> 2080</td><td> 2,74</td><td> 1,81</td><td> 2,51</td><td> 4,28</td>

<td>hsa-miR-497</td><td> 1995</td><td> 3,05</td><td> 3,11</td><td> 3,26</td><td> 0,58</td>

<td>hsa-miR-152</td><td> 2007</td><td> 7,72</td><td> 8,44</td><td> 6,59</td><td> 7,2</td>

<td>hsa-miR-181d</td><td> 2011</td><td> 8,56</td><td> 8,9</td><td> 7,83</td><td> 7,49</td>

<td>hsa-miR-660</td><td> 2144</td><td> 5,3</td><td> 5,36</td><td> 6,62</td><td> 6, 8</td>

<td>hsa-miR-52 6c</td><td> 2152</td><td> -0,58</td><td> -0,51</td><td> 2,42</td><td> 0,58</td>

<td>hsa-miR-584</td><td> 2176</td><td> 10,1</td><td> 10,43</td><td> 7, 6</td><td> 7,99</td>

<td>hsa-miR-299-3p</td><td> 2180</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-37 6a *</td><td> 2087</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-597</td><td> 2107</td><td> -0, 58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-511</td><td> 2109</td><td> 2,59</td><td> -0,01</td><td> 2,83</td><td> 2, 91</td>

<td>hsa-miR-599</td><td> 2113</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-141</td><td> 2117</td><td> -0,58</td><td> -0,51</td><td> 7,91</td><td> 8,21</td>

<td>hsa-miR-18b</td><td> 2125</td><td> 5,18</td><td> 5,41</td><td> 6,65</td><td> 6,98</td>

<td>hsa-miR-582</td><td> 2141</td><td> -0,58</td><td> -0,51</td><td> 4,9.</td><td> 4,87</td>

<td>hsa-miR-577</td><td> 2153</td><td> -0, 58</td><td> -0,51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-58 6</td><td> 2173</td><td> 2,11</td><td> 1,49</td><td> 2,47</td><td> 1,08</td>

<td>hsa-miR-380-3p</td><td> 2177</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-505</td><td> 2184</td><td> 5,06</td><td> 5, 45</td><td> 4,16</td><td> 4,58</td>

<td>hsa-miR-485-3p</td><td> 2196</td><td> 3,74</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-642</td><td> 2200</td><td> 4,22</td><td> 1,15</td><td> 2,42</td><td> 0,58</td>

<td>hsa-miR-615</td><td> 2204</td><td> 3, 94</td><td> -0,01</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-572</td><td> 2206</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-520d *</td><td> 2218</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-628</td><td> 2222</td><td> 3,59</td><td> 2,19</td><td> 2,17</td><td> 3,83</td>

<td>hsa-miR-518c *</td><td> 2226</td><td> 0,21</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-4 2 5-5p</td><td> 2234</td><td> , 8,86</td><td> 9,29</td><td> 9,01</td><td> 8,92</td>

<td>hsa-miR-432 *</td><td> 2266</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-661</td><td> 2274</td><td> 2,42</td><td> 1,81</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-421</td><td> 2185</td><td> .·.· 4,06</td><td> 5,49</td><td> . 6,41</td><td> 6, 43</td>

<td>hsa-miR-452 *</td><td> 2193</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-27b</td><td> 2303</td><td> 10,82</td><td> . 11,2</td><td> . 11,39</td><td> 11,55</td>

<td>hsa-miR-412</td><td> 2307</td><td> 2,59</td><td> -0,01</td><td> 0,51</td><td> 2,58</td>

<td>hsa-miR-579</td><td> 2311</td><td> -0,58</td><td> -0,01</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-519e *</td><td> 2315</td><td> -0,58</td><td> -0,51</td><td> 0, 51</td><td> 0,58</td>

<td>hsa-miR-588</td><td> 2327</td><td> 1,42</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-651</td><td> 2335</td><td> 1,71</td><td> 1,69</td><td> 3,39</td><td> 2,91</td>

<td>hsa-miR-557</td><td> 2339</td><td> 3,37</td><td> 2,49</td><td> 3,51</td><td> 0, 58</td>

<td>hsa-miR-507</td><td> 2343</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 3,39</td>

<td>hsa-miR-801</td><td> 2846</td><td> 5, 97</td><td> 3,08</td><td> &gt; 4,59</td><td> 1,88</td>

<td>hsa-miR-768-5p</td><td> 2854</td><td> 8,68</td><td> 8,01</td><td> 8,5</td><td> 8,38</td>

<td>hsa-miR-454-3p</td><td> 2858</td><td> 3</td><td> 3,37</td><td> 4,32</td><td> 3,78</td>

<td>hsa-miR-654</td><td> 2278</td><td> 2,22</td><td> -0,51</td><td> 1,31</td><td> 0,58</td>

<td>hsa-miR-4 98</td><td> 2298</td><td> 2,87</td><td> -0,51</td><td> 0,51</td><td> 2,67</td>

<td>hsa-miR-148b</td><td> 1362</td><td> 6, 83</td><td> 6,76</td><td> 6, 82</td><td> 6,69</td>

<td>hsa-miR-211</td><td> 1367</td><td> 3,56</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-127</td><td> 1377</td><td> 1,8</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-139</td><td> 1384</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-194</td><td> 1416</td><td> 8,57</td><td> 8,28</td><td> 4,64</td><td> 5,81</td>

<td>hsa-let-7e</td><td> 1421</td><td> 7, 42</td><td> 9,18</td><td> 8,74</td><td> 9,52</td>

<td>hsa-miR-345</td><td> 1444</td><td> &lt; 4,63</td><td> 4,62</td><td> 3, 68</td><td> 3,17</td>

<td>hsa-miR-1</td><td> 1448</td><td> -0,58</td><td> 0,29</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-30e-5p</td><td> 1461</td><td> -0, 58</td><td> -0, 51</td><td> 0,51</td><td> 0, 58</td>

<td>hsa-miR-134</td><td> 1470</td><td> -0,58</td><td> -0,51</td><td> 0,51</td><td> 0,58</td>

<td>hsa-miR-155</td><td> 1476</td><td> 8,21</td><td> 9,31</td><td> 4,32</td><td> 6,17</td>

<td>hsa-miR-374</td><td> 1480</td><td> 1,42</td><td> 1,79</td><td> 0,51</td><td> 1,38</td>

<td>hsa-miR-26b</td><td> 1484</td><td> 9,52</td><td> 10</td><td> 9,72</td><td> 10,43</td>

* The raw data were subtracted from the background, Log2 transformed and normalized.

The intensity of each oligo probe is based on the average of duplicated spots.

Data for all 4 67 human oligo probes are shown.

The normalized limit is calculated based on log2 (5 * non-spot background standard deviation + negative control probe signal with trim mean method).

normalized limit is calculated based on the 95^The percentile of the negative control probe signal.

A total of 236 Human Probes are Above the

Limit at least 1 sample.

A total of 158 Human Probes is Above TPT95 in at least 1 sample.

The Examples presented demonstrate the successful application of a cancer diagnostic assay and adverse pregnancy results with specificity, sensitivity and positive predictive values ​​greatly improved in relation to the currently available diagnoses, also providing other information on the stage, class and therapeutic response 15 that are not available in any other test format.

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