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Exome Sequencing Identified NRG3 as a Novel Susceptible Gene of Hirschsprung’s Disease in a Chinese Population

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Abstract

Hirschsprung’s disease (HSCR) is a complex developmental defect characterized by the absence of enteric ganglia in the gastrointestinal tract. Although the genetic defect of enteric nervous system (ENS) was identified to play a critical role in the progress of HSCR, the systemic genetic dissection of HSCR still needs to be clarified. In this study, we firstly performed exome sequencing of two HSCR patients from a Han Chinese family, including the affected mother and son. After the initial quality filtering (coverage ≥ 5X and SNP quality score ≥ 40) of the raw data, we identified 13,948 and 13,856 single nucleotide variants (SNVs), respectively. We subsequently compared the SNVs against public databases (dbSNP130, HapMap, and 1000 Genome Project) and obtained a total of 15 novel nonsynonymous SNVs in 15 genes, which were shared between these two patients. Follow-up Sanger sequencing and bioinformatics analysis highlighted variant c.853G>A (p.E285K) in NRG3, a gene involved in the development of ENS. In the validation phase, we sequenced all nine exons of NRG3 in 96 additional sporadic HSCR cases and 110 healthy individuals and identified another nonsynonymous variant c.1329G>A (p.M443I) and two synonymous variants c.828G>A (p.T276T) and c.1365T>A (p.P455P) only in the cases. Our results indicated that NRG3 may be a susceptibility gene for HSCR in a Chinese population.

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Acknowledgments

This work was supported by the Program for New Century Excellent Talents in University NCET-10-0388, Natural Science Foundation of Hubei 2010CDB03203 and 2011CDB304, and the Fundamental Research Funds for the Central Universities HUST: No. 2010QN005. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interest

The authors have declared no conflicts of interest.

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Authors and Affiliations

  1. Department of Pediatric Surgery, Union Hospital of Huazhong University of Science and Technology, Wuhan, 430022, China

    Jun Yang, Jiarui Pu, Jiahong Xia, Xiaoping Miao, Shaotao Tang & Guobin Wang

  2. Department of Epidemiology and Biostatistics and MOE Key Lab of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Shengyu Duan, Rong Zhong, Jieyun Yin, Juntao Ke, Xuzai Lu, Li Zou & Xiaoping Miao

  3. Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Hongmei Zhang & Anyuan Guo

  4. National Engineering Center for Biochip at Shanghai, No. 151 Libing Rd, Shanghai, 201203, China

    Zhidong Zhu & Huasheng Xiao

  5. Nextomics Biosciences Co., Ltd., Wuhan, 430074, China

    Depeng Wang

  6. Department of Pediatric Surgery, Wuhan Children’s Hospital, Wuhan, 430015, China

    Jun Yang

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  1. Jun Yang
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Correspondence to Xiaoping Miao, Shaotao Tang or Guobin Wang.

Additional information

Jun Yang, Shengyu Duan, and Rong Zhong contributed equally to this work.

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Supplementary Fig. 1

The pedigree of the affected Han Chinese family. Blue color represents HSCR patients. Patient II:3 is the mother and III:3 is the son (JPEG 23 kb)

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Supplementary Fig. 2

The sequence diagram of the identified false positive variants in UNC5C and C22orf42 (JPEG 50 kb)

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Supplementary Fig. 2b

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High resolution image (TIFF 326 kb)

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Yang, J., Duan, S., Zhong, R. et al. Exome Sequencing Identified NRG3 as a Novel Susceptible Gene of Hirschsprung’s Disease in a Chinese Population. Mol Neurobiol 47, 957–966 (2013). https://doi.org/10.1007/s12035-012-8392-4

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