Functional analysis across model systems implicates ribosomal proteins in growth and proliferation defects associated with hypoplastic left heart syndrome
Functional analysis across model systems implicates ribosomal proteins in growth and proliferation defects associated with hypoplastic left heart syndrome
Abstract Hypoplastic left heart syndrome (HLHS) is the most lethal congenital heart disease (CHD). The pathogenesis of HLHS is poorly understood and due to the likely oligogenic complexity of the disease, definitive HLHS-causing genes have not yet been identified. Postulating that impaired cardiomyocyte proliferation as a likely important contributing mechanism to HLHS pathogenesis, and we conducted a genome-wide siRNA screen to identify genes affecting proliferation of human iPSC-derived cardiomyocytes (hPSC-CMs). This yielded ribosomal protein (RP) genes as the most prominent class of effectors of CM proliferation. In parallel, whole genome sequencing and rare variant filtering of a cohort of 25 HLHS proband-parent trios with poor clinical outcome revealed enrichment of rare variants of RP genes. In addition, in another familial CHD case of HLHS proband we identified a rare, predicted-damaging promoter variant affecting RPS15A that was shared between the proband and a distant relative with CHD. Functional testing with an integrated multi-model system approach reinforced the idea that RP genes are major regulators of cardiac growth and proliferation, thus potentially contributing to hypoplastic phenotype observed in HLHS patients. Cardiac knockdown (KD) of RP genes with promoter or coding variants (RPS15A, RPS17, RPL26L1, RPL39, RPS15) reduced proliferation in generic hPSC-CMs and caused malformed hearts, heart-loss or even lethality in Drosophila. In zebrafish, diminished rps15a function caused reduced CM numbers, heart looping defects, or weakened contractility, while reduced rps17 or rpl39 function caused reduced ventricular size or systolic atrial dysfunction, respectively. Importantly, genetic interactions between RPS15A and core cardiac transcription factors TBX5 in CMs, Drosocross, pannier and tinman in flies, and tbx5 and nkx2-7 (nkx2-5 paralog) in fish, support a specific role for RP genes in heart development. Furthermore, RPS15A KD-induced heart/CM proliferation defects were significantly attenuated by p53 KD in both hPSC-CMs and zebrafish, and by Hippo activation (YAP/yorkie overexpression) in developing fly hearts. Based on these findings, we conclude that RP genes play critical roles in cardiogenesis and constitute an emerging novel class of gene candidates likely involved in HLHS and other CHDs.
Colas Alexandre R.、Missinato Maria A.、Zeng Xin-Xin I.、Berenguer Marie、Bodmer Rolf、Schroeder Analyne、Birker Katja、Olson Timothy M.、Ocorr Karen、Vogler Georg、Duester Greg、Kervadec Ana?s、Grossfeld Paul、Nelson Timothy J.、Nielsen Tanja、Walls Stanley M.、Lynott Michaela、Theis Jeanne L.
Development, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteCardiovascular Genetics Research, Mayo ClinicDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteUniversity of California San Diego, Rady Children?ˉs Hospital USACardiovascular Genetics Research, Mayo ClinicDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery Institute||Department of Biochemistry, Chemistry and Pharmacy, Freie Universit?t BerlinDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteDevelopment, Aging and Regeneration Program, Center for Genetic Disorders and Aging Research, Sanford-Burnham-Prebys Medical Discovery InstituteCardiovascular Genetics Research, Mayo Clinic
基础医学分子生物学生理学
Congenital heart diseaseHLHSwhole genome sequencingrare variant filteringcardiogenesisDrosophilaiPSCzebrafishRPS15ATP53yorkietinman/NKX2-5Dorsocross/TBX5pannier/GATA6
Colas Alexandre R.,Missinato Maria A.,Zeng Xin-Xin I.,Berenguer Marie,Bodmer Rolf,Schroeder Analyne,Birker Katja,Olson Timothy M.,Ocorr Karen,Vogler Georg,Duester Greg,Kervadec Ana?s,Grossfeld Paul,Nelson Timothy J.,Nielsen Tanja,Walls Stanley M.,Lynott Michaela,Theis Jeanne L..Functional analysis across model systems implicates ribosomal proteins in growth and proliferation defects associated with hypoplastic left heart syndrome[EB/OL].(2025-03-28)[2025-04-26].https://www.medrxiv.org/content/10.1101/2022.07.01.22277112.点此复制
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