Whole Exome Sequencing and Functional Characterization Increase Diagnostic Yield in Siblings with a 46, XY Difference of Sexual Development (DSD)

Overview

Journal J Steroid Biochem Mol Biol

Specialties Biochemistry
Molecular Biology

Date 2021 May 13

PMID 33984517

Authors

Sofia E Luna

Daniel J Wegner

Sarah Gale

Ping Yang

Abby Hollander

Lori St Dennis-Feezle

Zeina M Nabhan

Daniel S Ory

F Sessions Cole

Jennifer A Wambach

Affiliations

Soon will be listed here.

Abstract

Pathogenic biallelic variants in HSD17B3 result in 17β-hydroxysteroid dehydrogenase 3 (17β-HSD3) deficiency, variable disruption of testosterone production, and phenotypic diversity among 46, XY individuals with differences of sexual development (DSDs). We performed quad whole exome sequencing (WES) on two male siblings with microphallus, perineal hypospadias, and bifid scrotum and their unaffected parents. Both male siblings were compound heterozygous for a rare pathogenic HSD17B3 variant (c.239 G > A, p.R80Q) previously identified among individuals with 17β-HSD3 deficiency and a HSD17B3 variant (c.641A > G, p.E214 G) of uncertain significance. Following WES, the siblings underwent hCG stimulation testing with measurement of testosterone, androstenedione, and dihydrotestosterone which was non-diagnostic. To confirm pathogenicity of the HSD17B3 variants, we performed transient transfection of HEK-293 cells and measured conversion of radiolabeled androstenedione to testosterone. Both HSD17B3 variants decreased conversion of radiolabeled androstenedione to testosterone. As pathogenic HSD17B3 variants are rare causes of 46, XY DSD and hCG stimulation testing may not be diagnostic for 17β-HSD3 deficiency, WES in 46, XY individuals with DSDs can increase diagnostic yield and identify genomic variants for functional characterization of disruption of testosterone production.

References

Lee Y, Kirk J, Stanhope R, Johnston D, Harland S, Auchus R . Phenotypic variability in 17beta-hydroxysteroid dehydrogenase-3 deficiency and diagnostic pitfalls. Clin Endocrinol (Oxf). 2007; 67(1):20-8. DOI: 10.1111/j.1365-2265.2007.02829.x. View

Davydov E, Goode D, Sirota M, Cooper G, Sidow A, Batzoglou S . Identifying a high fraction of the human genome to be under selective constraint using GERP++. PLoS Comput Biol. 2010; 6(12):e1001025. PMC: 2996323. DOI: 10.1371/journal.pcbi.1001025. View

Saez J, de PERETTI E, Morera A, David M, Bertrand J . Familial male pseudohermaphroditism with gynecomastia due to a testicular 17-ketosteroid reductase defect. I. Studies in vivo. J Clin Endocrinol Metab. 1971; 32(5):604-10. DOI: 10.1210/jcem-32-5-604. View

Boehmer A, Brinkmann A, Sandkuijl L, Halley D, Niermeijer M, Andersson S . 17Beta-hydroxysteroid dehydrogenase-3 deficiency: diagnosis, phenotypic variability, population genetics, and worldwide distribution of ancient and de novo mutations. J Clin Endocrinol Metab. 1999; 84(12):4713-21. DOI: 10.1210/jcem.84.12.6174. View

Lek M, Karczewski K, Minikel E, Samocha K, Banks E, Fennell T . Analysis of protein-coding genetic variation in 60,706 humans. Nature. 2016; 536(7616):285-91. PMC: 5018207. DOI: 10.1038/nature19057. View

Ben Rhouma B, Kallabi F, Mahfoudh N, Ben Mahmoud A, Engeli R, Kamoun H . Novel cases of Tunisian patients with mutations in the gene encoding 17β-hydroxysteroid dehydrogenase type 3 and a founder effect. J Steroid Biochem Mol Biol. 2016; 165(Pt A):86-94. DOI: 10.1016/j.jsbmb.2016.03.007. View

McKeever B, Hawkins B, Geissler W, Wu L, Sheridan R, Mosley R . Amino acid substitution of arginine 80 in 17beta-hydroxysteroid dehydrogenase type 3 and its effect on NADPH cofactor binding and oxidation/reduction kinetics. Biochim Biophys Acta. 2002; 1601(1):29-37. DOI: 10.1016/s1570-9639(02)00434-x. View

Adzhubei I, Schmidt S, Peshkin L, Ramensky V, Gerasimova A, Bork P . A method and server for predicting damaging missense mutations. Nat Methods. 2010; 7(4):248-9. PMC: 2855889. DOI: 10.1038/nmeth0410-248. View

Yang Z, Ye L, Wang W, Zhao Y, Wang W, Jia H . 17β-Hydroxysteroid dehydrogenase 3 deficiency: Three case reports and a systematic review. J Steroid Biochem Mol Biol. 2017; 174:141-145. DOI: 10.1016/j.jsbmb.2017.08.012. View

10.

Moghrabi N, Hughes I, Dunaif A, Andersson S . Deleterious missense mutations and silent polymorphism in the human 17beta-hydroxysteroid dehydrogenase 3 gene (HSD17B3). J Clin Endocrinol Metab. 1998; 83(8):2855-60. DOI: 10.1210/jcem.83.8.5052. View

11.

Engeli R, Ben Rhouma B, Sager C, Tsachaki M, Birk J, Fakhfakh F . Biochemical analyses and molecular modeling explain the functional loss of 17β-hydroxysteroid dehydrogenase 3 mutant G133R in three Tunisian patients with 46, XY Disorders of Sex Development. J Steroid Biochem Mol Biol. 2015; 155(Pt A):147-54. DOI: 10.1016/j.jsbmb.2015.10.023. View

12.

Khattab A, Yuen T, Yau M, Domenice S, Costa E, Diya K . Pitfalls in hormonal diagnosis of 17-beta hydroxysteroid dehydrogenase III deficiency. J Pediatr Endocrinol Metab. 2014; 28(5-6):623-8. DOI: 10.1515/jpem-2014-0295. View

13.

Kolon T, Miller O . Comparison of single versus multiple dose regimens for the human chorionic gonadotropin stimulatory test. J Urol. 2001; 166(4):1451-4. View

14.

Chun S, Fay J . Identification of deleterious mutations within three human genomes. Genome Res. 2009; 19(9):1553-61. PMC: 2752137. DOI: 10.1101/gr.092619.109. View

15.

Andersson S, Geissler W, Wu L, Davis D, Grumbach M, New M . Molecular genetics and pathophysiology of 17 beta-hydroxysteroid dehydrogenase 3 deficiency. J Clin Endocrinol Metab. 1996; 81(1):130-6. DOI: 10.1210/jcem.81.1.8550739. View

16.

Camats N, Fluck C, Audi L . Oligogenic Origin of Differences of Sex Development in Humans. Int J Mol Sci. 2020; 21(5). PMC: 7084473. DOI: 10.3390/ijms21051809. View

17.

Wang K, Li M, Hakonarson H . ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res. 2010; 38(16):e164. PMC: 2938201. DOI: 10.1093/nar/gkq603. View

18.

George M, New M, Ten S, Sultan C, Bhangoo A . The clinical and molecular heterogeneity of 17βHSD-3 enzyme deficiency. Horm Res Paediatr. 2010; 74(4):229-240. DOI: 10.1159/000318004. View

19.

Cooper G, Stone E, Asimenos G, Green E, Batzoglou S, Sidow A . Distribution and intensity of constraint in mammalian genomic sequence. Genome Res. 2005; 15(7):901-13. PMC: 1172034. DOI: 10.1101/gr.3577405. View

20.

Andersson S, Moghrabi N . Physiology and molecular genetics of 17 beta-hydroxysteroid dehydrogenases. Steroids. 1997; 62(1):143-7. DOI: 10.1016/s0039-128x(96)00173-0. View