Single-cell RNA sequencing reveals that BMPR2 mutation regulates right ventricular function via ID genes

  • Eur Respir J. 2022 Jul 7;60(1):2100327. doi: 10.1183/13993003.00327-2021.
Mingxia Du  1  2 ,  Haibin Jiang  1  2 ,  Hongxian Liu  1  2 ,  Xin Zhao  1  2 ,  Yu Zhou  3 ,  Fang Zhou  2 ,  Chunmei Piao  4 ,  Guoqiang Xu  5 ,  Feng Ma  6 ,  Jianan Wang  7 ,  Frederic Perros  8 ,  Nicholas W Morrell  9 ,  Hong Gu  4 ,  Jun Yang  10
Affiliations
  • 1. Dept of Physiology, and Dept of Cardiology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
  • 2. Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China.
  • 3. Dept of General Intensive Care Unit, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
  • 4. Dept of Pediatric Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.
  • 5. Jiangsu Key Laboratory of Neuropsychiatric Diseases and College of Pharmaceutical Sciences, Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow University, Suzhou, China.
  • 6. Institute of Blood Transfusion, Chinese Academy of Medical Sciences, Chengdu, China.
  • 7. Dept of Cardiology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
  • 8. Université Paris-Saclay, AP-HP, INSERM UMR_S 999, Service de Pneumologie et Soins Intensifs Respiratoires, Hôpital de Bicêtre, Le Kremlin Bicêtre, France.
  • 9. Dept of Medicine, University of Cambridge School of Clinical Medicine, Addenbrooke's Hospital, Cambridge, UK.
  • 10. Dept of Physiology, and Dept of Cardiology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China [email protected].
PMID: 34857612 DOI: 10.1183/13993003.00327-2021
Abstract

Background: 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.

Methods: 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).

Results: 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.

Conclusion: 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.

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