US009719087B2 a2) United States Patent (0) Patent No.: US 9,719,087 B2 Olson et al. (45) Date of Patent: *Aug. 1, 2017 (54) MICRO-RNA FAMILY THAT MODULATES A61LK 39/3955 (2013.01); AGLK 45/06 FIBROSIS AND USES THEREOF (2013.01); A6IL 31/08 (2013.01); AGIL 31/16 (2013.01); C12N 9/16 (2013.01); C12N (71) Applicant: THE BOARD OF REGENTS, THE 15/8509 (2013.01); AOIK 2207/30 (2013.01); UNIVERSITY OF TEXAS SYSTEM, AOIK 2217/052 (2013.01); AOLK 2217/075 Austin, TX (US) (2013.01); AOIK 2217/15 (2013.01); AOIK 2217/206 (2013.01); AOIK 2227/105 (72) Inventors: Erie N. Olson, Dallas, TX (US); Eva (2013.01); AOIK 2267/0375 (2013.01); AIL van Rooij, Utrecht (NL) 2300/258 (2013.01); A6IL 2300/45 (2013.01); : AOIL 2420/06 (2013.01); C12N 2310/113 (73) Assignee: THE BOARD OF REGENTS, THE (2013.01); CI2N 2310/141 (2013.01); CI2N UNIVERSITY OF TEXAS SYSTEM, 2310/315 (2013.01); C12N 2310/321 Austin, TX (US) (2013.01); C12N 2310/346 (2013.01); C12N (*) Notice: Subjectto any disclaimer, the termbe this (013.01ars orb01301CDN US.C.patent154(b)is extendedby 0 ordays.adjusted under 2320/32 (2013.01);4 . CI2N 2330/10yor(2013.01) (58) Field of Classification Search This patent is subject to a terminal dis- CPC vieceeceseeseeeeeeee C12N 15/113; C12N 2310/141 claimer. See application file for complete search history. — (21) Appl. No.: 14/592,699 (56) References Cited (22) Filed: Jan. 8, 2015 U.S. PATENT DOCUMENTS (65) Prior Publication Data 7,232,806 B2 6/2007 Tuschlet al. US 2016/0145608 Al May 26, 2016 7,674,617 B2 3/2010 Kim etal. 8,440,636 B2* 5/2013 Olson wu... AOIK 67/0276 Related U.S. Application Data 514/44 A 8,940,711 B2 1/2015 Olsonet al. (60) Division of application No. 13/840,823, filed on Mar. 8,940,712 B2 1/2015 Olsonetal. 15, 2013, now Pat. No. 8,940,712, which is a (Continued) continuation of application No. 12/671,445, filed as application No. PCT/US2008/071839 on Jul. 31, FOREIGN PATENT DOCUMENTS 2008, now Pat. No. 8,440,636. EP 1440981 42 7/2004 (60) Provisional application No. 61/047,014,filed on Apr. EP 1627925 Al 2/2006 22, 2008, provisional application No. 60/980,303, (Continued) filed on Oct. 16, 2007, provisional application No. 60/952,917, filed on Jul. 31, 2007. OTHER PUBLICATIONS (51) Int. Cl. Van Rooij et al. “Control of Stress-Dependent Cardiac Growth and CIQN 15/113 (2010.01) Gene Expression by a MicroRNA”, Science, Apr. 27, 2007, vol. AOIK 67/027 (2006.01) 316, Issue 5824, p. 575-579. AGIK 38/22 (2006.01) (Continued) CI2N Y16 (2006.01) CI2N 15/85 (2006.01) AGIK 31/7105 (2006.01) Primary Examiner — J. E. Angell AGIK 31/712 (2006.01) AGIK 31/7125 (2006.01) AGIK 31/713 (2006.01) 6) ABSTRACT A6IK 39/395 (2006.01) The present invention relates to the identification of a AGIK 45/06 (2006.01) microRNA family, designated miR-29a-c, that is a key A6IK 31/07 (2006.01) regulator of fibrosis in cardiac tissue. The inventors show AGIK 31/355 (2006.01) that members of the miR-29 family are down-regulated in A6IK 31/375 (2006.01) the heart tissue in responseto stress, and are up-regulated in AGIL 31/08 (2006.01) heart tissue of mice that are resistant to both stress and AGIL 31/16 (2006.01) fibrosis. Also provided are methods of modulating expres- (52) U.S. Cl. sion and activity of the miR-29 family of miRNAs as a CPC we CI2N 15/113 (2013.01); AOLK 67/0276 treatmentfor fibrotic disease, including cardiac hypertrophy, (2013.01); A61K 31/07 (2013.01); A6IK skeletal muscle fibrosis other fibrosis related diseases and 31/355 (2013.01); A6IK 31/375 (2013.01); collagen loss-related disease. AGIK 31/712 (2013.01); A6IK 31/713 (2013.01); A61K 31/7105 (2013.01); A6IK 31/7125 (2013.01); A6LK 38/2242 (2013.01); 22 Claims, 29 Drawing Sheets US 9,719,087 B2 Page 2 (56) References Cited WO WO 2007/081720 A2 7/2007 WO WO 2007/081740 A2 7/2007 U.S. PATENT DOCUMENTS WO WO 2007/109236 A2 9/2007 WO WO 2007/112754 A2 10/2007 — 8,940,713 B2 1/2015 Olson et al. WO WO 2008/016924 A2 2/2008 9,376,681 B2 6/2016 Montgomery etal. WO WO 2008/043521 A2 4/2008 2005/0059005 Al 3/2005 Tuschl etal. WO WO 2008/061537 A2 5/2008 2005/0222399 Al 10/2005 Bentwich WO WO 2008/116267 Al 10/2008 — 2005/0261218 Al 11/2005 Esau etal. WO WO 2008/147839 Al 12/2008 — 2006/00 19286 Al 1/2006 Horvitz et al. WO WO 2009/012263 A2 1/2009 2006/0063730 Al 3/2006 Moniaetal. WO WO 2009/026576 Al 2/2009 2006/0105360 Al 5/2006 Croceet al. WO WO 2009/043353 A2 4/2009 2006/0185027 Al 8/2006 Bartel et al. WO WO 2009/058818 A2 5/2009 2006/0247 193 Al 11/2006 Taira et al. WO WO 2009/062169 A2 5/2009 2007/0087335 Al 4/2007 Brahmachariet al. WO WO 2009/111375 A2 9/2009 2007/0092882 Al 4/2007 Wangetal. WO WO 2009/114681 A2 9/2009 2008/0050722 Al 2/2008 Kim etal. WO WO 2009/121031 Al 10/2009 — 2008/0176766 Al 7/2008 Brownetal. WO WO 2009/149182 Al 12/2009 — 2009/0053718 Al 2/2009 Naguibnevaet al. WO WO 2010/036939 A2 4/2010 2009/0092980 Al 4/2009 Arenzet al. WO WO 2010/048585 A2 4/2010 2009/0143326 Al 6/2009 Obadetal. 2009/028 1167 Al 11/2009 Shenetal. OTHER PUBLICATIONS 2009/0286969 Al 11/2009 Esau etal. 2009/029 1906 Al 11/2009 Esau etal. Chenget al., “MicroRNAs Are Aberrantly Expressed. in Hypertro- 2009/029 1907 Al 11/2009 Esau etal. phic Heart,” Am. J. Pathol. 170(6):1831-1840 (2007). 2009/0293 148 Al 11/2009 Ren etal. Kim, “International Search Report,” 4 pages, from International 2009/03 17369 Al 12/2009 Hosodaetal. Patent Appl. No. PCT/US2008/071839, Korean Intellectual Prop- 2009/0326049 Al 12/2009 Aristarkhovetal. 2010/0029003 Al 2/2010 Bartel et al. erty Office, Daejon, Republic of Korea (mailed Jan. 6, 2009). Rooij et al., “A signature pattern of stress-responsive microRNAs 2010/00875 12 4/2010 Tuschl etal. that can evoke cardiac hypertrophy and heart failure,” Proc. Natl. 2010/00875 13 4/2010 Tuschl etal. 2010/0093837 4/2010 Tuschl etal. Acad. Sci. USA 103(48): 18255-18260 (2006). 2010/0099748 4/2010 Tuschl etal. Tatsuguchi et al., “Expression of MicroRNAs is Dynamically 2010/0113561 5/2010 Tuschl etal. Regulated During Cardiomyocyte Hypertrophy,” J. Mol. Cell. 2016/0145607 5/2016 Olson et al. Cardiol. 42(6):1137-1141 (2007). 2016/0145609 5/2016 Olson et al. Lagos-Quintanaet al., “New microRNAsfrom mouse and human,” 2016/0355815 12/2016 Montgomery etal. RNA,vol. 9:175-179, 2003. Lagos-Quintanaet al., “Identification of tissue-specific microRNAs FOREIGN PATENT DOCUMENTS from mouse,” Current Biology, vol. 12:735-739, 2002. Sempere et al., “Expression profiling of mammalian microRNAs EP 1777301 A2 4/2007 uncovers a subset of brain-expressed microRNAs with possible EP 1959012 A2 8/2008 roles in murine and human neuronal differentiation,” Genome EP 2113567 Al 11/2009 Biology, vol. 5:R13, 2004. JP 2006-5 19008 8/2006 Mott et al., “miR-29 regulates Mcl-1 protein expression and. JP 2006-292367 10/2006 apoptosis,” Oncogene, vol. 26: 6133-6140, 2007. WO WO 03/029459 A2 4/2003 Heet al., “Overexpression of Micro Ribonucleic Acid 29, Highly WO WO 2004/076622 A2 9/2004 Up-Regulated in Diabetic Rats, Leads to Insulin Resistance in WO WO 2005/013901 A2 2/2005 3T3-L1 Adipocytes,” Molecular Endocrinology, vol. 21: 2785- WO WO 2005/017145 Al 2/2005 2794, 2007. WO WO 2005/040419 Al 5/2005 Zhaoet al., “Dysregulation of Cardiogenesis, Cardiac Conduction, WO WO 2005/047505 A2 5/2005 WO WO 2005/056797 Al 6/2005 and Cell Cycle in Mice Lacking miRNA-1-2,” Cell, vol. 129: WO WO 2005/118806 A2 12/2005 303-317, 2007. WO WO 2006/033020 A2 3/2006 Van Rooij et al., “Dysregulation of microRNAsafter myocardial WO WO 2006/033928 A2 3/2006 infarction reveals a role of miR-29 in cardiac fibrosis,” Proc. Natl. WO WO 2006/081284 A2 8/2006 Acad. Sci. USA, vol. 105: 13027-13032, 2008. WO WO 2006/108473 Al 10/2006 Spindler, Supplementary European Search Report for European WO WO2006/111512 Al 10/2006 Application No. 08797006.7, 10 pages, European Patent Office, The WO WO 2006/128245 Al 12/2006 Hague, mailed Aug. 17, 2011. WO WO 2006/133022 A2 12/2006 Pekarsky Y. et al., “Tell expression in chronic lymphocytic leukemia WO WO 2006/137941 A2 12/2006 is regulated by miR-29 and miR-181,” Cancer Research, vol. WO WO 2007/000668 A2 1/2007 66:11590-11593, 2006. WO WO 2007/016548 A2 2/2007 WO WO 2007/033023 A2 3/2007 Written Opinion of the International Searching Authority mailed on WO WO 2007/042899 A2 4/2007 Jan. 6, 2009, for PCT Application No. PCT/US2008/071839, filed. WO WO 2007/067695 A2 6/2007 on Jul. 31, 2008, 5 pages. WO WO 2007/070483 A2 6/2007 WO WO 2007/081680 A2 7/2007 * cited by examiner B2 9,719,087 US t “Sig if DONNODYVYWWOSYpOVOTEAY JS QOS-HIW GIMBW (s :oN ar a5) 1.2 wou ¥ a.