Human Genetics

, Volume 137, Issue 3, pp 265–277 | Cite as

Next-generation sequencing reveals genetic landscape in 46, XY disorders of sexual development patients with variable phenotypes

  • Hao Wang
  • Lele Zhang
  • Nan Wang
  • Hui Zhu
  • Bing Han
  • Feng Sun
  • Haijun Yao
  • Qiang Zhang
  • Wenjiao Zhu
  • Tong Cheng
  • Kaixiang Cheng
  • Yang Liu
  • Shuangxia Zhao
  • Huaidong SongEmail author
  • Jie QiaoEmail author
Original Investigation


Disorders of sexual development (DSD) are rare congenital conditions in which chromosomal, gonadal, or anatomical sex is atypical. Currently, less than 20% of patients receive an accurate genetic diagnosis. Targeted next-generation sequencing, consisting of 33 candidate genes and 47 genes involved in sexual differentiation and development, was performed on 70 46, XY DSD patients. Functional assays were performed to evaluate the expression and transcriptional activity of one reported and nine novel mutations of NR5A1. In total, 113 mutations, including 86 novel and 27 reported sites in 40 genes, were identified in 52 patients. Among them, 37 mutations from 19 genes were first identified in 46, XY DSD patients, including EGF, LHX9, and CST9. Nine patients displayed biallelic mutations, 12 had mutations in sex chromosome genes and 14 had monoallelic mutations in NR5A1, BMP4, and WT1. Higher frequency mutations were identified in AR, SRD5A2, and NR5A1. Six missense, one frameshift, and one three-nucleotide deletion mutations of NR5A1 were shown to impair the transactivation ability with an altered nuclear aggregation of p.T29K and p.N44del variants. Multiple genetic mutations were identified in 33 of the 70 patients. The targeted sequencing panel provides an efficient method for the etiological diagnosis of 46, XY DSD patients and expands the candidate genes and inherited patterns.



We are grateful to the patients and their families for their supporting and participation in this study.

Author contributions

HW, LZ, and NW contributed equally to this work. HW, LZ, and FS performed the targeted exome sequencing and sequencing data analysis. Prediction of variants, verification by Sanger sequencing, and cell culture experiments was completed by HW and NW. BH completed protein structural analysis. HZ, BH, HY, WZ, TC, QZ, KC, and YL performed recruitment of patients, collected, and interpreted clinical data. SZ and HW did the statistical analysis. HS and JQ conceived and designed the experiments and revised the manuscript. All authors have seen and approved the final manuscript.


This work was supported by grants from the National Natural Science Foundation of China (Grant Number: 81570753, 81430019) and the Shanghai Committee for Science and Technology (Grant Number: 15ZR1425000).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

439_2018_1879_MOESM1_ESM.pdf (528 kb)
Supplementary material 1 (PDF 527 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Endocrinology, Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  2. 2.Research Centre for Clinical Medicine, Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  3. 3.Department of Urology, Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  4. 4.Department of Plastic Surgery, Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

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