The interplay of common genetic variants NRG1 rs2439302 and RET rs2435357 increases the risk of developing Hirschsprung's disease

Shuiqing Chi; Shuai Li; Guoqing Cao; Jialing Guo; Yunqiao Han; Yun Zhou; Xi Zhang; Yibo Li; Zhibin Luo; Xiangyang Li; Liying Rong; Mengxin Zhang; Linglu Li; Shaotao Tang

doi:10.3389/fcell.2023.1184799

The interplay of common genetic variants NRG1 rs2439302 and RET rs2435357 increases the risk of developing Hirschsprung's disease

Front Cell Dev Biol. 2023 Jul 7:11:1184799. doi: 10.3389/fcell.2023.1184799. eCollection 2023.

Authors

Shuiqing Chi¹, Shuai Li¹, Guoqing Cao¹, Jialing Guo¹, Yunqiao Han², Yun Zhou¹, Xi Zhang¹, Yibo Li¹, Zhibin Luo¹, Xiangyang Li¹, Liying Rong¹, Mengxin Zhang¹, Linglu Li³, Shaotao Tang¹

Affiliations

¹ Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
² Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
³ China Zebrafish Resource Center, National Aquatic Biological Resource Center, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China.

Abstract

Introduction: As a congenital and genetically related disease, many single nucleotide polymorphisms (SNPs) have been reported to be associated with the risk of HSCR. Our previous research showed that SNP rs2439302 (NRG1) interacted with rs2435357 (RET) to increase the risk of HSCR development. However, the underlying molecular mechanism is still not well understood. Methods: SNP rs2439302 (NRG1) and rs2435357 (RET) were genotyped in 470 HSCR cases. The expression of NRG1 and RET was investigated in the colon of HSCR patients. Knockdown of the NRG1 and RET homologs was performed in zebrafish to investigate their synergistic effect on ENS development. The effect of SNP rs2439302 and rs2435357 polymorphism on neuron proliferation, migration, and differentiation were investigated in SHSY-5Y cells and IPSCs. Results: Significant downregulation of NRG1 and RET expression was noticed in the aganglionic segment of HSCR patients and SHSY-5Y cells with rs2439302 GG/rs2435357 TT genotype. NRG1 and RET double mutants caused the most severe reduction in enteric neuron numbers than NRG1 single mutant or RET single mutant in the hindgut of zebrafish. SHSY-5Y cells and IPSCs with rs2439302 GG/rs2435357 TT genotype exhibited a decreased proliferative, migration, and differentiative capacity. CTCF showed a considerably higher binding ability to SNP rs2439302 CC than GG. NRG1 reduction caused a further decrease in SOX10 expression via the PI3K/Akt pathway, which regulates RET expression by directly binding to rs2435357. Discussion: SNP rs2439302 (NRG1) GG increases the risk of developing HSCR by affecting the binding of transcription factor CTCF and interacting with rs2435357 (RET) to regulate RET expression via the PI3K/Akt/SOX10 pathway.

Keywords: Hirschsprung’s disease; Nrg1; PI3K/AKT; RET; neuron development.

Grants and funding

This study was supported by the National Natural Science Foundations of China (Grant Nos. 81873848 and 81700497). The source of funding was not involved in study design, data collection, analysis, interpretation or paper writing.