Single-cell RNA sequencing reveals that BMPR2 mutation regulates right ventricular function via ID genes.
Single-cell RNA sequencing reveals that BMPR2 mutation regulates right ventricular function via ID genes.
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单细胞RNA测序揭示BMPR2突变通过ID基因调节右心室功能
DOI：
10.1183/13993003.00327-2021
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发表时间：
2022-07
影响因子：
24.3
通讯作者：
Yang, Jun
中科院分区：
文献类型：
--
作者：
Du, Mingxia;Jiang, Haibin;Liu, Hongxian;Zhao, Xin;Zhou, Yu;Zhou, Fang;Piao, Chunmei;Xu, Guoqiang;Ma, Feng;Wang, Jianan;Perros, Frederic;Morrell, Nicholas W.;Gu, Hong;Yang, Jun
骨形态发生蛋白II型受体(BMPR2)突变在先天性心脏病相关性肺动脉高压(CHD-PAH)患者中被发现。我们的研究旨在阐明BMPR2信号的缺陷是否通过心脏发育过程中DNA结合蛋白(ID)的下游效应物和抑制物参与了CHD患者PAH的进展。为了证实IDs是心脏中胚层前体细胞(CMPS)中BMPR2信号的下游效应者，并参与了PAH，我们建立了心肌细胞特异性ID1/3基因敲除小鼠(IDs CDKO)，25只小鼠中有12只发生了轻度PAH，血流动力学指标改变和肺血管重构。此外，我们还获得了ID1和ID3双敲除(IDsKO)人胚胎干细胞，它们概括了CHD-PAH诱导的多能干细胞(IPSCs)的BMPR2信号缺陷。共聚焦显微镜实验证明，从BMP受体突变的CHD-PAH患者的IPSCs分化而来的心肌细胞表现出心脏分化功能障碍和钙瞬变减少。CHD-PAH IPSCs和Bmpr2+/-大鼠右心室Smad1/5磷酸化和ID1、ID3表达降低。此外，超声波显示33%的IDs cDKO小鼠的室间隔和肺返流存在可检测到的缺陷。从小鼠右室分离的心肌细胞也显示出钙瞬变减少和肌节缩短。单细胞RNA测序分析显示，与野生型细胞相比，Ids KO细胞的CMPS分化受阻，USP9X表达下调。我们发现BMPR2信号通过IDs和USP9X来调节心脏分化，ID1和ID3的表达缺失是导致BMPR2突变的CHD-PAH患者心肌细胞功能障碍的原因之一。本研究首次报道了DNA结合蛋白基因敲除小鼠发生肺动脉高压，USP9X基因是BMPR2突变的CHD-PAH患者心脏发育过程中ID的下游效应因子。Https://bit.ly/3ciUNim
Mutations in bone morphogenetic protein type II receptor (BMPR2) have been found in patients with congenital heart disease-associated pulmonary arterial hypertension (CHD-PAH). Our study aimed to clarify whether deficient BMPR2 signalling acts through downstream effectors, inhibitors of DNA-binding proteins (IDs) during heart development to contribute to the progress of PAH in CHD patients. To confirm that IDs are downstream effectors of BMPR2 signalling in cardiac mesoderm progenitors (CMPs) and contribute to PAH, we generated cardiomyocyte-specific Id 1/3 knockout mice (Ids cDKO), and 12 out of 25 developed mild PAH with altered haemodynamic indices and pulmonary vascular remodelling. Moreover, we generated ID1 and ID3 double-knockout (IDs KO) human embryonic stem cells that recapitulated the BMPR2 signalling deficiency of CHD-PAH induced pluripotent stem cells (iPSCs). Cardiomyocytes differentiated from iPSCs derived from CHD-PAH patients with BMP receptor mutations exhibited dysfunctional cardiac differentiation and reduced calcium (Ca2+) transients, as evidenced by confocal microscopy experiments. Smad1/5 phosphorylation and ID1 and ID3 expression were reduced in CHD-PAH iPSCs and in Bmpr2+/– rat right ventricles. Moreover, ultrasound revealed that 33% of Ids cDKO mice had detectable defects in their ventricular septum and pulmonary regurgitation. Cardiomyocytes isolated from mouse right ventricles also showed reduced Ca2+ transients and shortened sarcomeres. Single-cell RNA sequencing analysis revealed impaired differentiation of CMPs and downregulated USP9X expression in IDs KO cells compared with wild-type cells. We found that BMPR2 signals through IDs and USP9X to regulate cardiac differentiation, and the loss of ID1 and ID3 expression contributes to cardiomyocyte dysfunction in CHD-PAH patients with BMPR2 mutations. This study reports for the first time that inhibitor of DNA-binding protein knockout mice developed pulmonary arterial hypertension and that the USP9X gene was a downstream effector of ID during heart development in CHD-PAH patients with BMPR2 mutations. https://bit.ly/3ciUNim
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影响因子：
2.7
作者：
Beppu, Hideyuki;Malhotra, Rajeev;Beppu, Yuko;Lepore, John J.;Parmacek, Michael S.;Bloch, Kenneth D.
通讯作者：
Bloch, Kenneth D.
影响因子：
10.5
作者：
Cunningham TJ;Yu MS;McKeithan WL;Spiering S;Carrette F;Huang CT;Bushway PJ;Tierney M;Albini S;Giacca M;Mano M;Puri PL;Sacco A;Ruiz-Lozano P;Riou JF;Umbhauer M;Duester G;Mercola M;Colas AR
通讯作者：
Colas AR
影响因子：
24.3
作者：
Roberts, KE;McElroy, JJ;Morse, JH
通讯作者：
Morse, JH
影响因子：
37.8
作者：
Tian, Wen;Jiang, Xinguo;Nicolls, Mark R.
通讯作者：
Nicolls, Mark R.
基于游离载体的 CRISPR/Cas9 系统，可在人类多能干细胞中高效进行基因敲除
影响因子：
4.6
作者：
Xie Y;Wang D;Lan F;Wei G;Ni T;Chai R;Liu D;Hu S;Li M;Li D;Wang H;Wang Y
通讯作者：
Wang Y