Josowitz Rebecca, Mulero-Navarro Sonia, Rodriguez Nelson A, Falce Christine, Cohen Ninette, Ullian Erik M, Weiss Lauren A, Rauen Katherine A, Sobie Eric A, Gelb Bruce D
The Mindich Child Health and Development Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Stem Cell Reports. 2016 Sep 13;7(3):355-369. doi: 10.1016/j.stemcr.2016.07.018. Epub 2016 Aug 25.
Germline mutations in BRAF cause cardio-facio-cutaneous syndrome (CFCS), whereby 40% of patients develop hypertrophic cardiomyopathy (HCM). As the role of the RAS/MAPK pathway in HCM pathogenesis is unclear, we generated a human induced pluripotent stem cell (hiPSC) model for CFCS from three patients with activating BRAF mutations. By cell sorting for SIRPα and CD90, we generated a method to examine hiPSC-derived cell type-specific phenotypes and cellular interactions underpinning HCM. BRAF-mutant SIRPα(+)/CD90(-) cardiomyocytes displayed cellular hypertrophy, pro-hypertrophic gene expression, and intrinsic calcium-handling defects. BRAF-mutant SIRPα(-)/CD90(+) cells, which were fibroblast-like, exhibited a pro-fibrotic phenotype and partially modulated cardiomyocyte hypertrophy through transforming growth factor β (TGFβ) paracrine signaling. Inhibition of TGFβ or RAS/MAPK signaling rescued the hypertrophic phenotype. Thus, cell autonomous and non-autonomous defects underlie HCM due to BRAF mutations. TGFβ inhibition may be a useful therapeutic option for patients with HCM due to RASopathies or other etiologies.
BRAF基因的种系突变会导致心脏-面部-皮肤综合征(CFCS),其中40%的患者会发展为肥厚型心肌病(HCM)。由于RAS/MAPK信号通路在HCM发病机制中的作用尚不清楚,我们从三名具有激活型BRAF突变的患者中构建了一个用于CFCS的人类诱导多能干细胞(hiPSC)模型。通过对信号调节蛋白α(SIRPα)和CD90进行细胞分选,我们开发了一种方法来检测hiPSC衍生的细胞类型特异性表型以及构成HCM基础的细胞间相互作用。携带BRAF突变的SIRPα(+)/CD90(-)心肌细胞表现出细胞肥大、促肥大基因表达以及内在钙处理缺陷。呈成纤维细胞样的携带BRAF突变的SIRPα(-)/CD90(+)细胞表现出促纤维化表型,并通过转化生长因子β(TGFβ)旁分泌信号部分调节心肌细胞肥大。抑制TGFβ或RAS/MAPK信号可挽救肥大表型。因此,细胞自主和非自主缺陷是BRAF突变导致HCM的基础。对于因RAS病或其他病因导致HCM的患者,抑制TGFβ可能是一种有效的治疗选择。