PMID- 31101927 OWN - NLM STAT- MEDLINE DCOM- 20200619 LR - 20220601 IS - 1432-1440 (Electronic) IS - 0946-2716 (Print) IS - 0946-2716 (Linking) VI - 97 IP - 7 DP - 2019 Jul TI - Therapeutic potential of AAV9-S15D-RLC gene delivery in humanized MYL2 mouse model of HCM. PG - 1033-1047 LID - 10.1007/s00109-019-01791-z [doi] AB - Familial hypertrophic cardiomyopathy (HCM) is an autosomal dominant disorder characterized by ventricular hypertrophy, myofibrillar disarray, and fibrosis, and is primarily caused by mutations in sarcomeric genes. With no definitive cure for HCM, there is an urgent need for the development of novel preventive and reparative therapies. This study is focused on aspartic acid-to-valine (D166V) mutation in the myosin regulatory light chain, RLC (MYL2 gene), associated with a malignant form of HCM. Since myosin RLC phosphorylation is critical for normal cardiac function, we aimed to exploit this post-translational modification via phosphomimetic-RLC gene therapy. We hypothesized that mimicking/modulating cardiac RLC phosphorylation in non-phosphorylatable D166V myocardium would improve heart function of HCM-D166V mice. Adeno-associated virus, serotype-9 (AAV9) was used to deliver phosphomimetic human RLC variant with serine-to-aspartic acid substitution at Ser15-RLC phosphorylation site (S15D-RLC) into the hearts of humanized HCM-D166V mice. Improvement of heart function was monitored by echocardiography, invasive hemodynamics (PV-loops) and muscle contractile mechanics. A significant increase in cardiac output and stroke work and a decrease in relaxation constant, Tau, shown to be prolonged in HCM mice, were observed in AAV- vs. PBS-injected HCM mice. Strain analysis showed enhanced myocardial longitudinal shortening in AAV-treated vs. control mice. In addition, increased maximal contractile force was observed in skinned papillary muscles from AAV-injected HCM hearts. Our data suggest that myosin RLC phosphorylation may have important translational implications for the treatment of RLC mutations-induced HCM and possibly play a role in other disease settings accompanied by depressed Ser15-RLC phosphorylation. KEY MESSAGES: HCM-D166V mice show decreased RLC phosphorylation and decompensated function. AAV9-S15D-RLC gene therapy in HCM-D166V mice, but not in WT-RLC, results in improved heart performance. Global longitudinal strain analysis shows enhanced contractility in AAV vs controls. Increased systolic and diastolic function is paralleled by higher contractile force. Phosphomimic S15D-RLC has a therapeutic potential for HCM. FAU - Yadav, Sunil AU - Yadav S AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Yuan, Chen-Ching AU - Yuan CC AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Kazmierczak, Katarzyna AU - Kazmierczak K AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Liang, Jingsheng AU - Liang J AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Huang, Wenrui AU - Huang W AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Takeuchi, Lauro M AU - Takeuchi LM AD - Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Kanashiro-Takeuchi, Rosemeire M AU - Kanashiro-Takeuchi RM AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. AD - Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. FAU - Szczesna-Cordary, Danuta AU - Szczesna-Cordary D AUID- ORCID: 0000-0002-8441-0722 AD - Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. dszczesna@med.miami.edu. LA - eng GR - R01 HL123255/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20190517 PL - Germany TA - J Mol Med (Berl) JT - Journal of molecular medicine (Berlin, Germany) JID - 9504370 RN - 0 (Myosin Light Chains) RN - 147336-22-9 (Green Fluorescent Proteins) RN - RMB44WO89X (Hydroxyproline) SB - IM MH - Animals MH - Cardiomyopathy, Hypertrophic, Familial/diagnostic imaging/genetics/physiopathology/*therapy MH - Dependovirus/*genetics MH - Disease Models, Animal MH - Echocardiography MH - Female MH - Fibrosis MH - *Gene Transfer Techniques MH - Genetic Vectors/*metabolism MH - Green Fluorescent Proteins/metabolism MH - HEK293 Cells MH - Hemodynamics MH - Humans MH - Hydroxyproline/metabolism MH - Male MH - Mice MH - Muscle Contraction MH - Muscles/metabolism MH - Myosin Light Chains/*genetics/*therapeutic use PMC - PMC6584042 MID - NIHMS1529612 OTO - NOTNLM OT - Adeno-associated virus OT - D166V-mutation OT - In vivo rescue of function OT - Regulatory light chain (RLC) OT - S15D-phosphorylation mimic EDAT- 2019/05/19 06:00 MHDA- 2020/06/20 06:00 PMCR- 2020/07/01 CRDT- 2019/05/19 06:00 PHST- 2018/11/14 00:00 [received] PHST- 2019/05/01 00:00 [accepted] PHST- 2019/04/24 00:00 [revised] PHST- 2019/05/19 06:00 [pubmed] PHST- 2020/06/20 06:00 [medline] PHST- 2019/05/19 06:00 [entrez] PHST- 2020/07/01 00:00 [pmc-release] AID - 10.1007/s00109-019-01791-z [pii] AID - 10.1007/s00109-019-01791-z [doi] PST - ppublish SO - J Mol Med (Berl). 2019 Jul;97(7):1033-1047. doi: 10.1007/s00109-019-01791-z. Epub 2019 May 17.