Silencing of CCR4-NOT complex subunits affects heart structure and function

Autor: Anaïs Kervadec, Karen Ocorr, Sreehari Kalvakuri, Rolf Bodmer, Santiago Pineda, Andrew A. Hicks, Claudia B. Volpato, Anthony Cammarato, Lisa Elmén, Peter P. Pramstaller, Nakissa N. Alayari, Alexandre R. Colas, Luisa Foco, Alessandra Rossini
Rok vydání: 2020
Předmět:
0301 basic medicine
Medicine (miscellaneous)
lcsh:Medicine
Action Potentials
030204 cardiovascular system & hematology
Long-QT syndrome
hiPSC
Animals
Genetically Modified
0302 clinical medicine
Immunology and Microbiology (miscellaneous)
Heart Rate
Drosophila heart
Gene expression
Morphogenesis
Drosophila Proteins
GWAS
Myocytes
Cardiac
Cardiomyocytes
Gene knockdown
Heart development
Intracellular Signaling Peptides and Proteins
Gene Expression Regulation
Developmental
RNA-Binding Proteins
Cell biology
Long QT Syndrome
Drosophila melanogaster
Arrhythmia
lcsh:RB1-214
Research Article
Induced Pluripotent Stem Cells
Neuroscience (miscellaneous)
Biology
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
Ribonucleases
CCR4-NOT complex
lcsh:Pathology
Gene silencing
Animals
Humans
Gene Silencing
Gene
Cell Proliferation
Messenger RNA
lcsh:R
Promoter
Dros
Repressor Proteins
030104 developmental biology
Exoribonucleases
CNOT1
Genome-Wide Association Study
HeLa Cells
Transcription Factors
Zdroj: Disease Models & Mechanisms
article-version (VoR) Version of Record
Disease Models & Mechanisms, Vol 13, Iss 7 (2020)
ISSN: 1754-8411
Popis: The identification of genetic variants that predispose individuals to cardiovascular disease and a better understanding of their targets would be highly advantageous. Genome-wide association studies have identified variants that associate with QT-interval length (a measure of myocardial repolarization). Three of the strongest associating variants (single-nucleotide polymorphisms) are located in the putative promotor region of CNOT1, a gene encoding the central CNOT1 subunit of CCR4-NOT: a multifunctional, conserved complex regulating gene expression and mRNA stability and turnover. We isolated the minimum fragment of the CNOT1 promoter containing all three variants from individuals homozygous for the QT risk alleles and demonstrated that the haplotype associating with longer QT interval caused reduced reporter expression in a cardiac cell line, suggesting that reduced CNOT1 expression might contribute to abnormal QT intervals. Systematic siRNA-mediated knockdown of CCR4-NOT components in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) revealed that silencing CNOT1 and other CCR4-NOT genes reduced their proliferative capacity. Silencing CNOT7 also shortened action potential duration. Furthermore, the cardiac-specific knockdown of Drosophila orthologs of CCR4-NOT genes in vivo (CNOT1/Not1 and CNOT7/8/Pop2) was either lethal or resulted in dilated cardiomyopathy, reduced contractility or a propensity for arrhythmia. Silencing CNOT2/Not2, CNOT4/Not4 and CNOT6/6L/twin also affected cardiac chamber size and contractility. Developmental studies suggested that CNOT1/Not1 and CNOT7/8/Pop2 are required during cardiac remodeling from larval to adult stages. To summarize, we have demonstrated how disease-associated genes identified by GWAS can be investigated by combining human cardiomyocyte cell-based and whole-organism in vivo heart models. Our results also suggest a potential link of CNOT1 and CNOT7/8 to QT alterations and further establish a crucial role of the CCR4-NOT complex in heart development and function. This article has an associated First Person interview with the first author of the paper.
Summary: Genome-wide association studies combined with in vitro human cardiac cell assays and a model organism suitable for heart studies in vivo connect CNOT1, CNOT7 and overall the CCR4-NOT complex to human heart disease and morbidity.
Databáze: OpenAIRE
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