Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function

Heart failure (HF) as a result of myocardial infarction (MI) is a major cause of fatality worldwide. However, the cause of cardiac dysfunction succeeding MI has not been elucidated at a sarcomeric level. Thus, studying the alterations within the sarcomere is necessary to gain insights on the fundame...

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Main Authors: Markandran, Kasturi, Yu, Haiyang, Song, Weihua, Lam, Do Thuy Uyen Ha, Madathummal, Mufeeda Changaramvally, Ferenczi, Michael Alan
Other Authors: Lee Kong Chian School of Medicine (LKCMedicine)
Format: Article
Language:English
Published: 2022
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Online Access:https://hdl.handle.net/10356/161395
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Institution: Nanyang Technological University
Language: English
id sg-ntu-dr.10356-161395
record_format dspace
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Medicine
Cardiac Regulatory Light Chain
Enzymes
spellingShingle Science::Medicine
Cardiac Regulatory Light Chain
Enzymes
Markandran, Kasturi
Yu, Haiyang
Song, Weihua
Lam, Do Thuy Uyen Ha
Madathummal, Mufeeda Changaramvally
Ferenczi, Michael Alan
Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
description Heart failure (HF) as a result of myocardial infarction (MI) is a major cause of fatality worldwide. However, the cause of cardiac dysfunction succeeding MI has not been elucidated at a sarcomeric level. Thus, studying the alterations within the sarcomere is necessary to gain insights on the fundamental mechansims leading to HF and potentially uncover appropriate therapeutic targets. Since existing research portrays regulatory light chains (RLC) to be mediators of cardiac muscle contraction in both human and animal models, its role was further explored In this study, a detailed characterisation of the physiological changes (i.e., isometric force, calcium sensitivity and sarcomeric protein phosphorylation) was assessed in an MI mouse model, between 2D (2 days) and 28D post-MI, and the changes were related to the phosphorylation status of RLCs. MI mouse models were created via complete ligation of left anterior descending (LAD) coronary artery. Left ventricular (LV) papillary muscles were isolated and permeabilised for isometric force and Ca2+ sensitivity measurement, while the LV myocardium was used to assay sarcomeric proteins' (RLC, troponin I (TnI) and myosin binding protein-C (MyBP-C)) phosphorylation levels and enzyme (myosin light chain kinase (MLCK), zipper interacting protein kinase (ZIPK) and myosin phosphatase target subunit 2 (MYPT2)) expression levels. Finally, the potential for improving the contractility of diseased cardiac papillary fibres via the enhancement of RLC phosphorylation levels was investigated by employing RLC exchange methods, in vitro. RLC phosphorylation and isometric force potentiation were enhanced in the compensatory phase and decreased in the decompensatory phase of HF failure progression, respectively. There was no significant time-lag between the changes in RLC phosphorylation and isometric force during HF progression, suggesting that changes in RLC phosphorylation immediately affect force generation. Additionally, the in vitro increase in RLC phosphorylation levels in 14D post-MI muscle segments (decompensatory stage) enhanced its force of isometric contraction, substantiating its potential in HF treatment. Longitudinal observation unveils potential mechanisms involving MyBP-C and key enzymes regulating RLC phosphorylation, such as MLCK and MYPT2 (subunit of MLCP), during HF progression. This study primarily demonstrates that RLC phosphorylation is a key sarcomeric protein modification modulating cardiac function. This substantiates the possibility of using RLCs and their associated enzymes to treat HF.
author2 Lee Kong Chian School of Medicine (LKCMedicine)
author_facet Lee Kong Chian School of Medicine (LKCMedicine)
Markandran, Kasturi
Yu, Haiyang
Song, Weihua
Lam, Do Thuy Uyen Ha
Madathummal, Mufeeda Changaramvally
Ferenczi, Michael Alan
format Article
author Markandran, Kasturi
Yu, Haiyang
Song, Weihua
Lam, Do Thuy Uyen Ha
Madathummal, Mufeeda Changaramvally
Ferenczi, Michael Alan
author_sort Markandran, Kasturi
title Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
title_short Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
title_full Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
title_fullStr Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
title_full_unstemmed Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
title_sort functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function
publishDate 2022
url https://hdl.handle.net/10356/161395
_version_ 1759855163489648640
spelling sg-ntu-dr.10356-1613952023-03-05T16:51:11Z Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function Markandran, Kasturi Yu, Haiyang Song, Weihua Lam, Do Thuy Uyen Ha Madathummal, Mufeeda Changaramvally Ferenczi, Michael Alan Lee Kong Chian School of Medicine (LKCMedicine) Genome Institute of Singapore Yong Loo Lin School of Medicine, National University of Singapore A*STAR Microscopy Platform—Electron Microscopy Science::Medicine Cardiac Regulatory Light Chain Enzymes Heart failure (HF) as a result of myocardial infarction (MI) is a major cause of fatality worldwide. However, the cause of cardiac dysfunction succeeding MI has not been elucidated at a sarcomeric level. Thus, studying the alterations within the sarcomere is necessary to gain insights on the fundamental mechansims leading to HF and potentially uncover appropriate therapeutic targets. Since existing research portrays regulatory light chains (RLC) to be mediators of cardiac muscle contraction in both human and animal models, its role was further explored In this study, a detailed characterisation of the physiological changes (i.e., isometric force, calcium sensitivity and sarcomeric protein phosphorylation) was assessed in an MI mouse model, between 2D (2 days) and 28D post-MI, and the changes were related to the phosphorylation status of RLCs. MI mouse models were created via complete ligation of left anterior descending (LAD) coronary artery. Left ventricular (LV) papillary muscles were isolated and permeabilised for isometric force and Ca2+ sensitivity measurement, while the LV myocardium was used to assay sarcomeric proteins' (RLC, troponin I (TnI) and myosin binding protein-C (MyBP-C)) phosphorylation levels and enzyme (myosin light chain kinase (MLCK), zipper interacting protein kinase (ZIPK) and myosin phosphatase target subunit 2 (MYPT2)) expression levels. Finally, the potential for improving the contractility of diseased cardiac papillary fibres via the enhancement of RLC phosphorylation levels was investigated by employing RLC exchange methods, in vitro. RLC phosphorylation and isometric force potentiation were enhanced in the compensatory phase and decreased in the decompensatory phase of HF failure progression, respectively. There was no significant time-lag between the changes in RLC phosphorylation and isometric force during HF progression, suggesting that changes in RLC phosphorylation immediately affect force generation. Additionally, the in vitro increase in RLC phosphorylation levels in 14D post-MI muscle segments (decompensatory stage) enhanced its force of isometric contraction, substantiating its potential in HF treatment. Longitudinal observation unveils potential mechanisms involving MyBP-C and key enzymes regulating RLC phosphorylation, such as MLCK and MYPT2 (subunit of MLCP), during HF progression. This study primarily demonstrates that RLC phosphorylation is a key sarcomeric protein modification modulating cardiac function. This substantiates the possibility of using RLCs and their associated enzymes to treat HF. Ministry of Education (MOE) Published version The work presented herein was supported by the Singapore Ministry of Education under its Academic Research Fund Tier 2 (Project No. MOE2016-T2-1-106). 2022-08-30T08:00:39Z 2022-08-30T08:00:39Z 2022 Journal Article Markandran, K., Yu, H., Song, W., Lam, D. T. U. H., Madathummal, M. C. & Ferenczi, M. A. (2022). Functional and molecular characterisation of heart failure progression in mice and the role of myosin regulatory light chains in the recovery of cardiac muscle function. International Journal of Molecular Sciences, 23(1), 88-. https://dx.doi.org/10.3390/ijms23010088 1661-6596 https://hdl.handle.net/10356/161395 10.3390/ijms23010088 35008512 2-s2.0-85122329844 1 23 88 en MOE2016-T2-1-106 International Journal of Molecular Sciences © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). application/pdf