Therapeutic potential of AAV9-S15D-RLC gene delivery in humanized MYL2 mouse model of HCM

Sunil Yadav, Chen Ching Yuan, Katarzyna Kazmierczak, Jingsheng Liang, Wenrui Huang, Lauro M. Takeuchi, Rosemeire Takeuchi, Danuta Szczesna-Cordary

Research output: Contribution to journalArticle

Abstract

Abstract: 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.

Original languageEnglish (US)
JournalJournal of Molecular Medicine
DOIs
StatePublished - Jan 1 2019

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Dependovirus
Hypertrophic Cardiomyopathy
Genes
Phosphorylation
Therapeutics
Myosins
Aspartic Acid
Genetic Therapy
Serogroup
Familial Hypertrophic Cardiomyopathy
Myosin Light Chains
Mutation
Papillary Muscles
Valine
Post Translational Protein Processing
Mechanics
Cardiac Output
Serine
Hypertrophy
Echocardiography

Keywords

  • Adeno-associated virus
  • D166V-mutation
  • In vivo rescue of function
  • Regulatory light chain (RLC)
  • S15D-phosphorylation mimic

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery
  • Genetics(clinical)

Cite this

Therapeutic potential of AAV9-S15D-RLC gene delivery in humanized MYL2 mouse model of HCM. / Yadav, Sunil; Yuan, Chen Ching; Kazmierczak, Katarzyna; Liang, Jingsheng; Huang, Wenrui; Takeuchi, Lauro M.; Takeuchi, Rosemeire; Szczesna-Cordary, Danuta.

In: Journal of Molecular Medicine, 01.01.2019.

Research output: Contribution to journalArticle

Yadav, Sunil ; Yuan, Chen Ching ; Kazmierczak, Katarzyna ; Liang, Jingsheng ; Huang, Wenrui ; Takeuchi, Lauro M. ; Takeuchi, Rosemeire ; Szczesna-Cordary, Danuta. / Therapeutic potential of AAV9-S15D-RLC gene delivery in humanized MYL2 mouse model of HCM. In: Journal of Molecular Medicine. 2019.
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abstract = "Abstract: 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.",
keywords = "Adeno-associated virus, D166V-mutation, In vivo rescue of function, Regulatory light chain (RLC), S15D-phosphorylation mimic",
author = "Sunil Yadav and Yuan, {Chen Ching} and Katarzyna Kazmierczak and Jingsheng Liang and Wenrui Huang and Takeuchi, {Lauro M.} and Rosemeire Takeuchi and Danuta Szczesna-Cordary",
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AU - Yuan, Chen Ching

AU - Kazmierczak, Katarzyna

AU - Liang, Jingsheng

AU - Huang, Wenrui

AU - Takeuchi, Lauro M.

AU - Takeuchi, Rosemeire

AU - Szczesna-Cordary, Danuta

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N2 - Abstract: 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.

AB - Abstract: 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.

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