PMID- 30848941 OWN - NLM STAT- MEDLINE DCOM- 20200518 LR - 20211204 IS - 1530-6860 (Electronic) IS - 0892-6638 (Print) IS - 0892-6638 (Linking) VI - 33 IP - 6 DP - 2019 Jun TI - Disruption of both ROCK1 and ROCK2 genes in cardiomyocytes promotes autophagy and reduces cardiac fibrosis during aging. PG - 7348-7362 LID - 10.1096/fj.201802510R [doi] AB - In this study, we investigated the pathophysiological impact of Rho-associated coiled-coil-containing protein kinase (ROCK)1 and ROCK2 double deletion vs. single deletion on cardiac remodeling. Utilizing a cardiomyocyte-specific and tamoxifen-inducible MerCreMer recombinase (MCM), 3 mouse lines (MCM/ROCK1(fl/fl)/ROCK2(fl/fl), MCM/ROCK1(fl/fl), and MCM/ROCK2(fl/fl)) were generated. As early as 5 d after inducible deletion, the double ROCK knockout hearts exhibited reduced phosphorylation of myosin light chain (MLC) and focal adhesion kinase (FAK), supporting a role for ROCK activity in regulating the nonsarcomeric cytoskeleton. Moreover, the autophagy marker microtubule-associated proteins 1A-1B light chain 3B was increased in the double ROCK knockout, and these early molecular features persisted throughout aging. Mechanistically, the double ROCK knockout promoted age-associated or starvation-induced autophagy concomitant with reduced protein kinase B (AKT), mammalian target of rapamycin (mTOR), Unc-51-like kinase signaling, and cardiac fibrosis. In contrast, ROCK2 knockout hearts showed increased phosphorylated (p)-MLC and p-FAK levels, which were mostly attributable to a compensatory ROCK1 overactivation. Autophagy was inhibited at the baseline accompanying increased mTOR activity, leading to increased cardiac fibrosis in the ROCK2 knockout hearts. Finally, the loss of ROCK1 had no significant effect on p-MLC and p-FAK levels, mTOR signaling, or autophagy at baseline. In summary, deletions of ROCK isoforms in cardiomyocytes have different, even opposite, effects on endogenous ROCK activity and the MLC/FAK/AKT/mTOR signaling pathway, which is involved in autophagy and fibrosis of the heart.-Shi, J., Surma, M., Yang, Y., Wei, L. Disruption of both ROCK1 and ROCK2 genes in cardiomyocytes promotes autophagy and reduces cardiac fibrosis during aging. FAU - Shi, Jianjian AU - Shi J AD - Herman B. Wells Center for Pediatric Research, Department of Pediatrics, School of Medicine, Indiana University, Indianapolis, Indiana, USA. FAU - Surma, Michelle AU - Surma M AD - Herman B. Wells Center for Pediatric Research, Department of Pediatrics, School of Medicine, Indiana University, Indianapolis, Indiana, USA. FAU - Yang, Yang AU - Yang Y AD - Herman B. Wells Center for Pediatric Research, Department of Pediatrics, School of Medicine, Indiana University, Indianapolis, Indiana, USA. AD - Department of Cardiovascular Surgery, Xiangya Hospital, Central South University School of Medicine, Changsha, China; and. FAU - Wei, Lei AU - Wei L AD - Herman B. Wells Center for Pediatric Research, Department of Pediatrics, School of Medicine, Indiana University, Indianapolis, Indiana, USA. AD - Department of Cellular and Integrative Physiology, School of Medicine, Indiana University, Indianapolis, Indiana, USA. LA - eng GR - P01 HL085098/HL/NHLBI NIH HHS/United States GR - P01 HL134599/HL/NHLBI NIH HHS/United States GR - R01 HL107537/HL/NHLBI NIH HHS/United States PT - Comparative Study PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20190308 PL - United States TA - FASEB J JT - FASEB journal : official publication of the Federation of American Societies for Experimental Biology JID - 8804484 RN - 0 (Recombinant Proteins) RN - 094ZI81Y45 (Tamoxifen) RN - EC 2.7.1.1 (mTOR protein, mouse) RN - EC 2.7.11.1 (Rock1 protein, mouse) RN - EC 2.7.11.1 (Rock2 protein, mouse) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.1 (rho-Associated Kinases) SB - IM MH - Aging/genetics/metabolism/*pathology MH - Animals MH - Autophagy/genetics/*physiology MH - Crosses, Genetic MH - Enzyme Induction/drug effects MH - Female MH - Fibrosis MH - Gene Expression Regulation MH - Male MH - Mice MH - Mice, Knockout MH - Mice, Transgenic MH - Myocytes, Cardiac/*metabolism/pathology MH - Recombinant Proteins/biosynthesis MH - TOR Serine-Threonine Kinases/physiology MH - Tamoxifen/pharmacology MH - rho-Associated Kinases/deficiency/genetics/*physiology PMC - PMC6529334 OTO - NOTNLM OT - Rho kinase OT - cytoskeleton OT - mTOR COIS- The authors thank the Transgenic and Knock-Out Mouse Core, the Small Animal Ultrasound Core, and the Electron Microscopy Center at the Indiana University School of Medicine for assistance in the generation of Rho-associated coiled-coil-containing protein kinase (ROCK1)(fl/fl) and ROCK2(fl/fl) mice, echocardiography imaging, and electron microscopy imaging. This work was supported by U.S. National Institutes of Health, National Heart, Lung, and Blood Institute Grants HL107537 and HL134599; a Grant-in-Aid award from the American Heart Association, Midwest Affiliate (12GRNT12060525); and the Riley Children's Foundation (to L.W.). It was also supported by a Biomedical Research Grant from the Indiana University School of Medicine (2286128ZJ to J.S.) The authors declare no conflicts of interest. EDAT- 2019/03/09 06:00 MHDA- 2020/05/19 06:00 PMCR- 2020/03/08 CRDT- 2019/03/09 06:00 PHST- 2020/03/08 00:00 [pmc-release] PHST- 2019/03/09 06:00 [pubmed] PHST- 2020/05/19 06:00 [medline] PHST- 2019/03/09 06:00 [entrez] AID - FJ_201802510R [pii] AID - 10.1096/fj.201802510R [doi] PST - ppublish SO - FASEB J. 2019 Jun;33(6):7348-7362. doi: 10.1096/fj.201802510R. Epub 2019 Mar 8.