Abstract
Purpose: The study aimed to identify novel genes for idiopathic hypogonadotropic hypogonadism (IHH). Methods: A cohort of 1387 probands with IHH underwent exome sequencing and de novo, familial, and cohort-wide investigations. Functional studies were performed on 2 p190 Rho GTPase–activating proteins (p190 RhoGAP), ARHGAP35 and ARHGAP5, which involved in vivo modeling in larval zebrafish and an in vitro p190A-GAP activity assay. Results: Rare protein-truncating variants (PTVs; n = 5) and missense variants in the RhoGAP domain (n = 7) in ARHGAP35 were identified in IHH cases (rare variant enrichment: PTV [unadjusted P = 3.1E-06] and missense [adjusted P = 4.9E-03] vs controls). Zebrafish modeling using gnrh3:egfp phenotype assessment showed that mutant larvae with deficient arhgap35a, the predominant ARHGAP35 paralog in the zebrafish brain, display decreased GnRH3-GFP+ neuronal area, a readout for IHH. In vitro GAP activity studies showed that 1 rare missense variant [ARHGAP35 p.(Arg1284Trp)] had decreased GAP activity. Rare PTVs (n = 2) also were discovered in ARHGAP5, a paralog of ARHGAP35; however, arhgap5 zebrafish mutants did not display significant GnRH3-GFP+ abnormalities. Conclusion: This study identified ARHGAP35 as a new autosomal dominant genetic driver for IHH and ARHGAP5 as a candidate gene for IHH. These observations suggest a novel role for the p190 RhoGAP proteins in GnRH neuronal development and integrity.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2501-2515 |
| Number of pages | 15 |
| Journal | Genetics in Medicine |
| Volume | 24 |
| Issue number | 12 |
| DOIs | |
| State | Published - Dec 2022 |
Funding
We are grateful to the families with idiopathic hypogonadotropic hypogonadism for their support and willingness to participate in our research studies. We thank all providers who referred individuals for genetic research. We acknowledge Maxime Bouchard, McGill University, Montreal, Canada, for providing the pGEX-6P1 encoding GST-tagged GAP domain of p190A. We acknowledge Rebecca A. Rojas, Vanessa P.F. Lopes, and Nicole P. DiOrio for data curation. This research was made possible through access to the data and findings generated by the 100,000 Genomes Project (proband 14). The 100,000 Genomes Project is managed by Genomics England Limited (a wholly-owned company of the Department of Health and Social Care). The 100,000 Genomes Project is funded by the National Institute for Health Research and NHS England. The Wellcome Trust, Cancer Research UK, and the Medical Research Council have also funded research infrastructure. The 100,000 Genomes Project uses data provided by patients and collected by the National Health Service as part of their care and support. E.E.D. is the Ann Marie and Francis Klocke, MD Research Scholar. This work was supported by the US National Institutes of Health grants P50HD028138 (S.B.S., W.F.C., and R.B.) and P50HD104224 (S.B.S., M.F.L., and R.B.), Natural Sciences and Engineering Research Council of Canada grant RGPIN/04809-2017 (N.L.-V.), and the NIHR Oxford Biomedical Research Centre (A.O.M.W.). M.F.L. is supported by 5K23HD097296. R.B. is supported by R01HD096324. G.J.F. was supported by the Crusader Internship Fund from The College of the Holy Cross. R.S.T. was supported by an MRC DTP Studentship. The 100,000 Genomes Project and associated infrastructure are funded by the National Institute for Health Research and NHS England, the Wellcome Trust, Cancer Research UK, and the Medical Research Council. We are grateful to the families with idiopathic hypogonadotropic hypogonadism for their support and willingness to participate in our research studies. We thank all providers who referred individuals for genetic research. We acknowledge Maxime Bouchard, McGill University, Montreal, Canada, for providing the pGEX-6P1 encoding GST-tagged GAP domain of p190A. We acknowledge Rebecca A. Rojas, Vanessa P.F. Lopes, and Nicole P. DiOrio for data curation. This research was made possible through access to the data and findings generated by the 100,000 Genomes Project (proband 14). The 100,000 Genomes Project is managed by Genomics England Limited (a wholly-owned company of the Department of Health and Social Care). The 100,000 Genomes Project is funded by the National Institute for Health Research and NHS England . The Wellcome Trust, Cancer Research UK, and the Medical Research Council have also funded research infrastructure. The 100 , 000 Genomes Project uses data provided by patients and collected by the National Health Service as part of their care and support. E.E.D. is the Ann Marie and Francis Klocke, MD Research Scholar. This work was supported by the US National Institutes of Health grants P50HD028138 (S.B.S., W.F.C., and R.B.) and P50HD104224 (S.B.S., M.F.L., and R.B.), Natural Sciences and Engineering Research Council of Canada grant RGPIN/04809-2017 (N.L.-V.), and the NIHR Oxford Biomedical Research Centre (A.O.M.W.). M.F.L. is supported by 5K23HD097296. R.B. is supported by R01HD096324. G.J.F. was supported by the Crusader Internship Fund from The College of the Holy Cross . R.S.T. was supported by an MRC DTP Studentship . The 100,000 Genomes Project and associated infrastructure are funded by the National Institute for Health Research and NHS England , the Wellcome Trust, Cancer Research UK, and the Medical Research Council.
Keywords
- Developmental disorder
- Idiopathic hypogonadotropic hypogonadism
- Intellectual disability
- Puberty
- Rho GTPase–activating protein
ASJC Scopus subject areas
- Genetics(clinical)