COMPOSITIONS AND METHODS RELATED TO MIRNA MODULATION OF NEOVASCULARIZATION OR ANGIOGENESIS

• US 20090281167A1
• Filed: 05/08/2009
• Published: 11/12/2009
• Est. Priority Date: 05/08/2008
• Status: Active Grant

• Current Assignee: Johns Hopkins University, Mirna Therapeutics, Inc. (Synlogic, Inc.)
• Original Assignee: Mirna Therapeutics, Inc. (Synlogic, Inc.)
• Inventors: Campochiaro, Peter, Shen, Jikui, Brown, David, Kelnar, Kevin, Shelton, Jeffrey
• Granted Patent: US 8,258,111 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2525/207 siRNA, miRNA
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/118 Prognosis of disease development
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA

Abstract

The present invention concerns methods and compositions for diagnosing and/or treating vascular diseases including cancer, cardiac diseases, vascular diseases of the eye, and inflammatory diseases. The methods involve measuring the levels of one or multiple miRNAs in patient samples and using the test results to diagnose and/or predict an optimal treatment regimen for the patient. Compositions described in the invention include nucleic acids that function as miRNAs or miRNA inhibitors that can be introduced to a patient to reduce or increase vascularization as needed.

Claims

1. A method for reducing vascularization in a subject or tissue comprising administering to the subject or tissue in need of such a reduction, in an amount sufficient to reduce vascularization, one or more nucleic acid molecule comprising:

(a) a nucleic acid sequence that is at least 80% identical to one or more of miR-184, miR-31, miR-150, miR-409, miR-375, miR-129-5p, miR-142a, miR-29a, miR-129-3p, miR-10b, miR-96, miR-183, miR-16, miR-182, miR-191, miR-29c, miR-181c, miR-335, miR-7026, miR-210, miR-512-3p, miR-132, miR-500, miR-339, miR-511, miR-26b, miR-30b, or miR-15a, or complement thereof; and

/or

(b) an inhibitor of miR-451, miR-424, miR-146, miR-214, miR-199a, miR-181, miR-350, miR-21, miR-218, miR-148b, miR-106a, miR-205, miR-365, miR-299-5p, ambi-miR-7079, miR-200a, miR-351, miR-329, miR-122a, miR-20a, miR-520h, miR-142-5p, miR-203, miR-211, miR-145, let-7b, miR-93, miR-192, miR-201, miR-18a, miR-17-5p, miR-7085, miR-106b, or miR-223.

2. The method of claim 1, wherein the nucleic acid is administered topically, enterally, parenterally or intravitreally.

3. The method of claim 1, wherein the nucleic acid molecule is an RNA.

4. The method of claim 3, wherein the RNA comprises a complementary RNA region.

5. The method of claim 4, wherein the RNA comprises a hairpin structure.

6. The method of claim 4, wherein the RNA comprises two RNA strands.

7. The method of claim 3, wherein the RNA molecule includes a nucleotide analog or a modified nucleotide.

8. The method of claim 1, wherein the nucleic acid molecule or miRNA inhibitor is a DNA molecule or is produced from a DNA molecule.

9. The method of claim 8, wherein the miRNA inhibitor is an antisense oligonucleotide.

10. The method of claim 9, wherein the oligonucleotide comprises a nucleotide analog.

11. The method of claim 8, wherein the DNA is comprised in an expression cassette.

12. The method of claim 11, wherein the expression cassette is comprised in a plasmid expression vector.

13. The method of claim 11, wherein the nucleic acid molecule is comprised in a viral expression vector.

14. The method of claim 1, wherein the nucleic acid molecule is comprised in a lipid or viral delivery vehicle.

15. The method of claim 1, wherein the subject has, is at risk of developing, or is suspected of having ocular or retinal/choroidal neovascular diseases, cancer, diabetic nephropathy, rheumatoid arthritis, atherosclerotic plaques, endometriosis, Crohn'"'"'s disease, uterine fibroids, benign prostatic hyperplasia, or psoriasis.

16. A method of stimulating vascularization in subject or tissue comprising administering to a subject in need of such stimulation, in an amount sufficient to stimulate vascularization, one or more nucleic acid molecule comprising

(a) a miRNA sequence that is at least 80% identical to one or more of miR-451, miR-424, miR-146, miR-214, miR-199a, miR-181, miR-350, miR-21, miR-218, miR-148b, miR-106a, miR-205, miR-365, miR-299-5p, ambi-miR-7079, miR-200a, miR-351, miR-329, miR-122a, miR-20a, miR-520h, miR-142-5p, miR-203, miR-211, miR-145, let-7b, miR-93, miR-192, miR-201, miR-18a, miR-17-5p, miR-7085, miR-106b, and/or miR-223;

and/or

(b) an inhibitor of miR-184, miR-31, miR-150, miR-409, miR-375, miR-129-5p, miR-142a, miR-29a, miR-129-3p, miR-10b, miR-96, miR-183, miR-16, miR-182, miR-191, miR-29c, miR-181c, miR-335, miR-7026, miR-210, miR-512-3p, miR-132, miR-500, miR-339, miR-511, miR-26b, miR-30b, and/or miR-15a.

17. The method of claim 16, wherein the nucleic acid is administered topically, enterally, parenterally or intravitreally.

18. The method of claim 16 wherein the subject has, is at risk of developing, or is suspected of having coronary artery disease (CAD), cardiac failure, tissue injury, or ischemia.

19. (canceled)

20. (canceled)

21. A method for evaluating a patient comprising the steps of:

(a) determining expression levels of one or more of miR-451, miR-424, miR-146, miR-214, miR-199a, miR-181, miR-350, miR-21, miR-218, miR-148b, miR-106a, miR-205, miR-365, miR-299-5p, ambi-miR-7079, miR-200a, miR-351, miR-329, miR-122a, miR-20a, miR-520h, miR-142-5p, miR-203, miR-211, miR-145, let-7b, miR-93, miR-192, miR-201, miR-18a, miR-17-5p, miR-7085, miR-106b, miR-223, miR-184, miR-31, miR-150, miR-409, miR-375, miR-129-5p, miR-142a, miR-29a, miR-129-3p, miR-10b, miR-96, miR-183, miR-16, miR-182, miR-191, miR-29c, miR-181c, miR-335, miR-7026, miR-210, miR-512-3p, miR-132, miR-500, miR-339, miR-511, miR-26b, miR-30b, and/or miR-15a in a biological sample comprising a portion of a tissue or fluid associated with a condition associated with aberrant vacularization, and

(b) determining a diagnosis or prognosis for the aberrant vascularization condition based on the miRNA expression levels.

23. The method of claim 21, wherein determining a diagnosis is screening for a pathological condition, staging a pathological condition, or assessing response of a pathological condition to therapy.

22. (canceled)

24-44. -44. (canceled)