RT Journal Article
SR Electronic
T1 Alteration of myocardial structure and function in RAF1-associated Noonan syndrome: Insights from cardiac disease modeling based on patient-derived iPSCs
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.01.22.477319
DO 10.1101/2022.01.22.477319
A1 Saeideh Nakhaei-Rad
A1 Farhad Bazgir
A1 Julia Dahlmann
A1 Alexandra Viktoria Busley
A1 Marcel Buchholzer
A1 Fereshteh Haghighi
A1 Anne Schänzer
A1 Andreas Hahn
A1 Sebastian Kötter
A1 Denny Schanze
A1 Ruchika Anand
A1 Florian Funk
A1 Andrea Borchardt
A1 Annette Vera Kronenbitter
A1 Jürgen Scheller
A1 Roland P. Piekorz
A1 Andreas S. Reichert
A1 Marianne Volleth
A1 Matthew J. Wolf
A1 Ion Cristian Cirstea
A1 Bruce D. Gelb
A1 Marco Tartaglia
A1 Joachim Schmitt
A1 Martina Krüger
A1 Ingo Kutschka
A1 Lukas Cyganek
A1 Martin Zenker
A1 George Kensah
A1 Mohammad R. Ahmadian
YR 2022
UL http://biorxiv.org/content/early/2022/01/23/2022.01.22.477319.abstract
AB Noonan syndrome (NS), the most common among the RASopathies, is caused by germline variants in genes encoding components of the RAS-MAPK pathway. Distinct variants, including the recurrent Ser257Leu substitution in RAF1, are associated with severe hypertrophic cardiomyopathy (HCM). Here, we investigated the elusive mechanistic link between NS-associated RAF1S257L and HCM using three-dimensional cardiac bodies and bioartificial cardiac tissues generated from patient-derived induced pluripotent stem cells (iPSCs) harboring the pathogenic RAF1 c.770C>T missense change. We characterize the molecular, structural and functional consequences of aberrant RAF1 –associated signaling on the cardiac models. Ultrastructural assessment of the sarcomere revealed a shortening of the I-bands along the Z disc area in both iPSC-derived RAF1S257L cardiomyocytes, and myocardial tissue biopsies. The disease phenotype was partly reverted by using both MEK inhibition, and a gene-corrected isogenic RAF1L257S cell line. Collectively, our findings uncovered a direct link between a RASopathy gene variant and the abnormal sarcomere structure resulting in a cardiac dysfunction that remarkably recapitulates the human disease. These insights represent a basis to develop future targeted therapeutic approaches.Competing Interest StatementThe authors have declared no competing interest.ACadenylyl cyclaseACTN1actinin alpha 1APFalpha fetoproteinα-SMAalpha smooth muscle actinBNPbrain natriuretic peptideCACNA1Ccalcium voltage-gated channel subunit alpha1 CCBcardiac bodyCTcardiac tissueCMcardiomyocytescTNTcardiac troponin TDUSP1dual-specificity phosphataseEBembryoid bodyEBNA1Epstein-Barr nuclear antigen 1EMelectron microscopyERKextracellular regulated kinaseFOXOA2forkhead box A2GSK3βglycogen synthase kinase 3 betaHCMhypertrophic cardiomyopathyHDAChistone deacetylaseHPRT1hypoxanthine-guanine phosphoribosyltransferaseLTCCL-type calcium channelsMAPKmitogen-activated protein kinaseMEF2myocyte enhancer factor 2MEKMAP/ERK kinaseMEKiMAP/ERK kinase inhibitorMYHmyosin heavy chainMYLmyosin light chain 7iPSCinduced pluripotent stem cellsNFATnuclear factor of activated T-cellsNCnocodazoleNKX2.5NK2 homeobox 5NPPBnatriuretic peptide BNSNoonan syndromeOCT-4octamer-binding transcription factor 4PEphenylephrinep-H3phosphohistone 3PLNphospholambanRAFrapidly accelerated fibrosarcomaRT-PCRreverse transcriptase polymerase chainRyR2ryanodine receptor type-2SERCA2Asarco/endoplasmic reticulum Ca2+-ATPaseSOX2SRY-box transcription factor 2TNNC1troponin C1TNNI3troponin I3, cardiac typeTRP53transformation related protein 53TUBB3tubulin beta 3 class IIIWTwild-type.