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US008940712B2 a2) United States Patent (0) Patent No.: US 8,940,712 B2 Olsonet al. (45) Date of Patent: Jan. 27, 2015 (54) MICRO-RNA FAMILY THAT MODULATES 2310/141 (2013.01); CI2N 2310/315 (2013.01); FIBROSIS AND USES THEREOF C1I2N 2310/321 (2013.01); C12N 2310/346 (2013.01); CI2N 2310/3515 (2013.01); C12N (71) Applicant: The Board of Regents, The University 2320/31 (2013.01); C12N 2330/10 (2013.01) of Texas System,Austin, TX (US) USPC. ecsscssssssseesstvesnssnssresnesvesveecee 514/44 A (72) Inventors: Eric N. Olson, Dallas, TX (US); Eva (58) Field of Classification Search van Rooij, Utrecht (NL) USPC ooceecccccsscsecseeseeseeseeseeseeseesseeeeeeesneseees 514/44 A See application file for complete search history. (73) Assignee: The Board of Regents, The University of Texas System, Austin, TX (US) (56) References Cited (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 U.S. PATENT DOCUMENTS U.S.C. 154(6)by I day. 7,232,806 B2 6/2007 Tuschlet al. 7,674,617 B2 3/2010 Kimetal. (21) Appl. No.: 13/840,823 2005/0059005 Al 3/2005 Tuschlet al. 2005/0222399 Al 10/2005 Bentwich (22) Filed: Mar. 15, 2013 2005/0261218 Al 11/2005 Esau etal. 2006/0019286 Al 1/2006 Horvitz et al. (65) Prior Publication Data 2006/0105360 Al 5/2006 Croceet al. 2006/0185027 Al 8/2006 Bartelet al. US 2013/0195891 Al Aug. 1, 2013 2006/0247193 Al 11/2006 Taira etal. 2007/0087335 Al 4/2007 Brahmachari etal. 2007/0092882 Al 4/2007 Wangetal. 2008/0176766 Al 7/2008 Brownetal. (63) Continuation of application No. 12/671,445, filed as 2009/0053718 Al 2/2009 Naguibnevaetal. application No. PCT/US2008/071839 on Jul. 31, sooOaaaoe ‘i eons One "a 2008, now Pat. No. 8,440,636. 2009/0281167 Al 11/2009 Shenad.et&al. (60) Provisional application No. 61/047,014, filed on Apr. (Continued) 22, 2008, provisional application No. 60/980,303, filed on Oct. 16, 2007, provisional application No. FOREIGN PATENT DOCUMENTS 60/952,917,filed on Jul. 31, 2007. EP 1440981 A2 7/2004 (51) Int. Cl. EP 1627925 Al 2/2006 CI2N 15/11 (2006.01) (Continued) A6I1K 31/713 (2006.01) AOIK 67/027 (2006.01) OTHER PUBLICATIONS A61K 38/22 (2006.01) CI2N 9/16 (2006.01) Cheng et al., “MicroRNAs Are Aberrantly Expressed in CIQN 15/113 (2010.01) Hypertrophic Heart,’ Am. J. Pathol. 170(6):1831-1840 (2007). CI2QN 15/85 (2006.01) (Continued) AG6IK 31/7105 (2006.01) A6I1K 31/712 (2006.01) AG6IK 31/7125 2006.01 AGIK 39/395 (3006 01) Primary Examiner — J. E. Angell AG6IK 45/06 (2006.01) (74) Altorney, Agent, or Firm — Cooley LLP A61K 31/07 (2006.01) A6IK 31/355 (2006.01) A6I1K 31/375 (2006.01) (67) ABSTRACT (52) U.S. Cl. The present invention relates to the identification of a CPC wee A61K 31/713 (2013.01); AOLK 67/0276 microRNA family, designated miR-29a-c, that is a key regu- (2013.01); A6LK 38/2242 (2013.01); C12N lator of fibrosis in cardiac tissue. The inventors show that 9/16 (2013.01); CI2N 15/113 (2013.01); CI2N members of the miR-29 family are down-regulated in the 15/8509 (2013.01); AGLK 31/7105 (2013.01); heart tissue in responseto stress, and are up-regulatedin heart A61LK 31/712 (2013.01); A6LK 31/7125 tissue of mice that are resistant to both stress and fibrosis. (2013.01); A6LK 39/3955 (2013.01); A61K Also provided are methods of modulating expression and 45/06 (2013.01); A61K 31/07 (2013.01); A6LK activity of the miR-29 family of miRNAsas a treatment for 31/355 (2013.01); A6LK 31/375 (2013.01); fibrotic disease, including cardiac hypertrophy, skeletal AOLK 2207/30 (2013.01); AOLK 2217/052 muscle fibrosis other fibrosis related diseases and collagen (2013.01); AOLK 2217/075 (2013.01); AOILK loss-related disease. 2217/15 (2013.01); AOLK 2217/206 (2013.01); AOLK 2227/105 (2013.01); AOLK 2267/0375 (2013.01); CI2N 2310/113 (2013.01); C12N 16 Claims, 25 Drawing Sheets US 8,940,712 B2 Page 2 (56) References Cited WO WO2008/061537 A2 5/2008 Wo WO 2008/116267 Al 10/2008 U.S. PATENT DOCUMENTS Wo WO 2008/147839 Al 12/2008 — Wo WO 2009/012263 A2 1/2009 2009/0286969 Al 11/2009 Esauetal. WO WO 2009/026576 Al 2/2009 2009/0291906 Al 11/2009 Esauetal. WO WO 2009/043353 A2 4/2009 2009/0291907 Al 11/2009 Esauetal. WO WO 2009/058818 A2 5/2009 2009/0293148 Al 11/2009 Renetal. WO WO 2009/062169 A2 5/2009 2009/0317369 Al 12/2009 Hosodaet al. WO WO 2009/111375 A2 9/2009 2009/0326049 Al 12/2009 Aristarkhovetal. WO WO 2009/114681 A2 9/2009 2010/0029003 Al 2/2010 Bartel et al. WO WO 2009/121031 Al 10/2009 2010/0087512 Al 4/2010 Tuschlet al. we we sebseoso SS ot 2010/0087513 Al 4/2010 Tuschletal. 2010/0093837 Al 4/2010 Tuschl etal. WO WO 2010/048585 A2 4/2010 2010/0099748 Al 4/2010 Tuschlet al. OTHER PUBLICATIONS 2010/0113561 Al 5/2010 Tuschlet al. Kim, “International Search Report,” 4 pages, from International FOREIGN PATENT DOCUMENTS Patent Appl. No. PCT/US2008/071839,Korean Intellectual Property Office, Daejon, Republic of Korea (mailed Jan. 6, 2009). EP 1777301 A2 4/2007 Rooij et al., “A signature pattern of stress-responsive microRNAsthat EP 1959012 A2 8/2008 can evoke cardiac hypertrophy and heart failure,” Proc. Natl. Acad. EP 2113567 Al 11/2009 Sci. USA 103(48):18255-18260 (2006). JP 2006-5 19008 8/2006 Tatsuguchiet al., “Expression of MicroRNAsis Dynamically Regu- wi0 Wowuloooas9 AQ iOooos lated During Cardiomyocyte Hypertrophy,” J. Mol. Cell. Cardiol. woWo WO 2005/0139012004/076622 A2 3/20059/2004 Po)pasesQuintana8) altoeCon microictoRNAss fifrom mouse and. dhhuman,” wo wo aN40iio ‘i a8 Lagos-Quintanaet al., “Identification of tissue-specific microRNAs wo WO 2005/047505 A2 5/2005 from mouse,” Current Biology, vol. 12:735-739, 2002. ; wo WO 2005/056797 Al 6/2005 Sempere et al., “Expression profiling of mammalian microRNAs Wo WO 2005/118806 A2 12/2005 uncoversa subset ofbrain-expressed microRNAs with possible roles wo WO 2006/033020 A2 3/2006 in murine and human neuronal differentiation”’ Genome Biology, WO WO 2006/033928 A2 3/2006 vol. 5:R13, 2004. — wo WO 2006/081284 A2 8/2006 Mott et al., “miR-29 regulates Mcl-1 protein expression and WO WO 2006/108473 Al 10/2006 apoptosis,” Oncogene, vol. 26: 6133-6 140, 2007. WO WO 2006/111512 Al 10/2006 Heet al., “Overexpression of Micro Ribonucleic Acid 29, Highly WO WO 2006/128245 Al 12/2006 Up-Regulated in Diabetic Rats, Leads to Insulin Resistance in 3T3- wo WO 2006/133022 A2 12/2006 L1 Adipocytes,” Molecular Endocrinology, vol. 21: 2785-2794, Wo WO 2006/137941 A2 12/2006 2007. — WO WO 2007/000668 A2 1/2007 Zhaoet al., “Dysregulation of Cardiogenesis, Cardiac Conduction, WO WO 2007/016548 A2 2/2007 and Cell Cycle in Mice Lacking miRNA-1-2,” Cell, vol. 129: 303- — Wo WO 2007/033023 A2 3/2007 317, 2007. WO WO 2007/042899 A2 4/2007 Van Rooij et al., “Dysregulation of microRNAsafter myocardial wo wo sooooags “ ene infarction reveals a role of miR-29 in cardiac fibrosis,” Proc. Natl. Wo WO 2007/081680 A2 7/9007 Acad. Sci, USA, vol. 105: 13027-13032, 2008. wo WO 2007/081720 A2 7/2007 Spindler, Supplementary European Search Report for European Wo WO 2007/081740 A? 7/2007 Application No. 08797006.7, 10 pages, European Patent Office, The WO WO 2007/109236 A2 9/2007 Hague, mailed Aug. 17, 2011. ~~ Wo WO 2007/112754 A2 10/2007 Pekarsky Y. et al., “Tell expression in chronic lymphocytic leukemia WO WO 2008/016924 A2 2/2008 is regulated by miR-29 and miR-181,” Cancer Research, vol. Wo WO 2008/043521 A2 4/2008 66:11590-11593, 2006. 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