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Micro-RNA Biomarkers and Methods of Using Same

a microrna biomarker and biomarker technology, applied in the field of microrna biomarkers and methods of using same, can solve the problem of low healing rate of non-small cell lung carcinoma (nsclc)

Inactive Publication Date: 2012-09-06
BIOMIRNA HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0025]A further aim of the invention is to make available procedures which can be easily set up in laboratories, by analyzing biological samples collected from an individual.
[0029]A final aim of the invention is to provide pharmaceutical compounds and / or treatments for reducing and / or eliminating the risk of developing a pulmonary tumour.
[0100]In a sixty-sixth aspect in accordance with any one of aspects from the 60th to the 64th, the compound comprises a therapeutically effective quantity of the inhibitor of the expression of at least one of the miRNAs listed in Table Ia, Ic, IIa and / or Table IIc and / or Table Va, Vc, VIa and / or Table Vic, optionally all those listed in Table Ib, Id, IIb and / or Table IId, and / or Table Vb, Vd, VIb and / or Table VId, the quantity being able, for the over-expressed miRNAs with respect to the respective control level of expression, to reduce the values of the level of expression to the respective control level of expression.

Problems solved by technology

Notwithstanding recent progress in treatment of pulmonary cancer after resection and the use of specific treatments for determined molecular targets, the rate of healing of non-small-cell lung carcinoma (NSCLC) remains low due to the reappearance thereof in patients that are resistant to drugs or who present metastasis.

Method used

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Examples

Experimental program
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example 1

Studies, Materials & Methods

[0226]The present invention investigated the expression profile of miRNA in the plasma of individuals enrolled in screening protocols using spiral CT. This investigation was done with the aim of verifying the capability of miRNAs as a new class of biomolecular markers for: prediction of the risk of developing a tumour, in particular a pulmonary tumour, and diagnosis of the tumour, in particular pulmonary tumour, and thus as a prognostic aid for discriminating patients with indolent or aggressive pulmonary lesions.

[0227]Plasma samples taken from smoker individuals were used, where the individuals were over 50 years old, in a time parameter of between one and two years before detection with CT spiral of the presence of a pulmonary tumour in the same individuals. Also used were samples of plasma collected at the moment of the appearance of the disease (detected using spiral CT). The plasma samples were obtained from patients who had developed a pulmonary tum...

example 2

miRNA Treatment

[0313]The instant example demonstrates modifying the level of two microRNAs of our plasma signatures in a lung cancer cell line (A549). Mir-486 and mir-660 were down-modulated in plasma samples of patients with lung cancer and in particular in those who have developed the aggressive form of the disease. In FIG. 9 microRNA levels were measured by qReal-Time PCR in 20 paired tumor and normal lung tissue of the same patients enrolled in the CT-screening trial used as validation set. Row Ct data were normalized on the housekeeping miRNA RNU6B (DCt). The final expression values were obtained with the formula: 2̂(−DCt of the tumor tissue) / 2̂(−DCt of the normal lung). Values >1→upregulated in tumor tissue. Values<1→downregulated in tumor tissue. The results in FIG. 9 show that these two miRNA were downregulated in the tumor tissue compared with the normal lung tissue.

[0314]In FIG. 10 mirVana™ miRNA Mimic (Applied biosystem) were used to transfect lung cancer cell line expres...

example 3

Lung Cancer Detection and Survival

[0317]INT-IEO cohort (training set). Lung cancer was diagnosed in 38 subjects, 22 in the first 2 y and 16 from the 3rd to 5th y of screening, including one interval cancer at 4th y. The frequency of stage I was 63% (77% in first 2 y vs. 44% in the last 3 y), and adenocarcinoma was 71% (95% in first 2 y vs. 63% in the last 3 y; Table XI).

TABLE XICT YearCharacteristic1-23-5TotalLung Cancer221638Resected21 (95)12 (75)33* (87) Stage I17 (77) 7 (44)24 (63)Stage II-IV 5 (23) 9 (56)14 (37)Adeno17 (95)10 (63)27 (71)*28 tumor tissue and 24 normal lung samples were available for miRNA expression analysis. The number in parenthesis is the percent of all detected lung cancers.

[0318]Median follow-up time for the 38 lung cancer cases was 75 mo, with 60% 5-y overall survival (95% C.I.: 43-74%). Five-y overall survival was 92% for stage 1 and 7% for stages II-IV (P2 test, P=0.034). In the subset of CT year 1-2 / stage I, 5-y survival was 94% (95% C.I.: 65.0-99.1). In...

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Abstract

A procedure and an apparatus are described for identifying individuals at risk of pulmonary tumour and / or for diagnosing a pulmonary tumour using the study of levels of expression of miRNA in the blood or another biological fluid. Also described are a method and a compound for reducing or eliminating a risk of pulmonary tumour by rebalancing the miRNAs that are underexpressed or overexpressed.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of Italian Patent Application Nos. MI2011A000172, MI2011A000173 and MI2011A000174, each filed Feb. 7, 2011 and U.S. Provisional Application No. 61 / 522,328, filed Aug. 11, 2011. The contents of each of these applications are incorporated herein by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention concerns methods for identifying and using, in pre-diagnostic and / or diagnostic stages, special molecular bio-markers identifiable in biological samples, such as for example whole blood, serum, plasma, saliva or bronchia condensate collected from an individual.[0003]In more detail, the invention relates to methods for identifying individuals at risk of tumour, in particular pulmonary tumour. The invention also concerns methods for determining a presence and / or level of aggressiveness of a tumour, for example a pulmonary tumour, in an individual.[0004]The invention also relates to diag...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K31/7088A61P35/00C40B60/12C40B30/04C40B40/06
CPCC12Q1/6886C12Q2600/118C12Q2600/178C12Q2600/158A61P3/00A61P35/00G16B30/00
Inventor SOZZI, GABRIELLAPASTORINO, UGOBOERI, MATTIA
Owner BIOMIRNA HLDG
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