Novel Heterozygous Genetic Variants in Patients with 46,XY Gonadal Dysgenesis

Vasundhera Chauhan; Viveka P. Jyotsna; Vandana Jain; Rajesh Khadgawat; Rima Dada

doi:10.1055/s-0042-114778

Hormone and Metabolic Research, Inhaltsverzeichnis

Horm Metab Res 2017; 49(01): 36-42
DOI: 10.1055/s-0042-114778

Endocrine Care

Novel Heterozygous Genetic Variants in Patients with 46,XY Gonadal Dysgenesis

Vasundhera Chauhan

¹Department of Endocrinology and Metabolism, All India Institute of Medical Sciences, New Delhi, India

,

Viveka P. Jyotsna

¹Department of Endocrinology and Metabolism, All India Institute of Medical Sciences, New Delhi, India

,

Vandana Jain

²Division of Paediatric Endocrinology, Department of Paediatrics, All India Institute of Medical Sciences, New Delhi, India

,

Rajesh Khadgawat

¹Department of Endocrinology and Metabolism, All India Institute of Medical Sciences, New Delhi, India

,

Rima Dada

³Department of Anatomy, All India Institute of Medical Sciences, New Delhi, India

› Institutsangaben

Abstract

46,XY gonadal dysgenesis (GD) constitutes a rare group of disorders characterized by the presence of dysfunctional testes in genotypic males. The molecular etiology is not known in about 2 thirds of instances. The aim of this study was to identify the genetic cause in patients with 46,XY gonadal dysgenesis. Based on clinical, cytogenetic, and biochemical screening, 10 patients with 46,XY GD were recruited. Direct sequencing of SRY, NR5A1, SOX9, DAX1, DHH, DMRT1 genes was carried out for molecular analysis. Among 10 patients, 5 were diagnosed with complete gonadal dysgenesis (CGD), 3 with partial gonadal dysgenesis (PGD), and 3 with testicular agenesis. Molecular analysis revealed 12 heterozygous genetic changes, 4 of which were novel. One (c.416T>A) was observed in evolutionary conserved region of DMRT1 gene in a patient with CGD and was found to be probably damaging on in silico analysis. Other 3 were identified in NR5A1 gene (c.990+22 C>A, c.1387+1403T>A and p.131P), but their association with gonadal dysgenesis is not evident from our study. These genetic changes were absent in parents and 50 healthy control samples, which were also studied. With targeted sequencing approach, a molecular diagnosis was made in only one patient with 46,XY GD. The application of new genomic technologies is required for the precise evaluation of these rare genetic defects.

Key words

46,XY gonadal dysgenesis - genes - mutation - molecular diagnosis

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Referenzen

References
1 Eggers S, Sinclair A. Mammalian sex determination – insights from humans and mice. Chromosome Res 2012; 20: 215-238
2 Lee PA, Houk CP, Ahmed SF, Hughes IA. International Consensus Conference on Intersex organized by the Lawson Wilkins Pediatric Endocrine Society and the European Society for Paediatric Endocrinology . Consensus statement on management of intersex disorders. International Consensus Conference on Intersex. Pediatrics 2006; 118: e488-e500
3 Sinclair AH, Berta P, Palmer MS, Hawkins JR, Griffiths BL, Smith MJ, Foster JW, Frischauf AM, Lovell-Badge R, Goodfellow PN. A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif. Nature 1990; 346: 240-244
4 Röpke A, Tewes A-C, Gromoll J, Kliesch S, Wieacker P, Tüttelmann F. Comprehensive sequence analysis of the NR5A1 gene encoding steroidogenic factor 1 in a large group of infertile males. Eur J Hum Genet 2013; 21: 1012-1015
5 Tremblay JJ, Viger RS. A mutated form of steroidogenic factor 1 (SF-1 G35E) that causes sex reversal in humans fails to synergize with transcription factor GATA-4. J Biol Chem 2003; 278: 42637-42642
6 Sekido R, Lovell-Badge R. Sex determination involves synergistic action of SRY and SF1 on a specific Sox9 enhancer. Nature 2008; 453: 930-934
7 Foster JW, Dominguez-Steglich MA, Guioli S, Kwok C, Weller PA, Stevanović M, Weissenbach J, Mansour S, Young ID, Goodfellow PN. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 1994; 372: 525-530
8 Iyer AK, McCabe ERB. Molecular mechanisms of DAX1 action. Mol Genet Metab 2004; 83: 60-73
9 Landau Z, Hanukoglu A, Sack J, Goldstein N, Weintrob N, Eliakim A, Gillis D, Sagi M, Shomrat R, Kosinovsky EB, Anikster Y. Clinical and genetic heterogeneity of congenital adrenal hypoplasia due to NR0B1 gene mutations. Clin Endocrinol (Oxf) 2010; 72: 448-454
10 Bardoni B, Zanaria E, Guioli S, Floridia G, Worley KC, Tonini G, Ferrante E, Chiumello G, McCabe ER, Fraccaro M. A dosage sensitive locus at chromosome Xp21 is involved in male to female sex reversal. Nat Genet 1994; 7: 497-501
11 de Mello MP, Coeli FB, Assumpção JG, Castro TM, Maciel-Guerra AT, Marques-de-Faria AP, Baptista MTM, Guerra-Júnior G. Novel DMRT1 3′UTR+11insT mutation associated to XY partial gonadal dysgenesis. Arq Bras Endocrinol Metabol 2010; 54: 749-753
12 Matson CK, Murphy MW, Sarver AL, Griswold MD, Bardwell VJ, Zarkower D. DMRT1 prevents female reprogramming in the postnatal mammalian testis. Nature 2011; 476: 101-104
13 Baetens D, Mladenov W, Delle Chiaie B, Menten B, Desloovere A, Iotova V, Callewaert B, Van Laecke E, Hoebeke P, De Baere E, Cools M. Extensive clinical, hormonal and genetic screening in a large consecutive series of 46,XY neonates and infants with atypical sexual development. Orphanet J Rare Dis 2014; 9: 209
14 Ahmed SF, Khwaja O, Hughes IA. The role of a clinical score in the assessment of ambiguous genitalia. BJU Int 2000; 85: 120-124
15 Davenport M, Brain C, Vandenberg C, Zappala S, Duffy P, Ransley PG, Grant D. The use of the hCG stimulation test in the endocrine evaluation of cryptorchidism. Br J Urol 1995; 76: 790-794
16 Knorr D, Beckmann D, Bidlingmaier F, Helmig FJ, Sippell WG. Plasma testosterone in male puberty. II. hCG stimulation test in boys with hypospadia. Acta Endocrinol (Copenh.) 1979; 90: 365-371
17 Georg I, Bagheri-Fam S, Knower KC, Wieacker P, Scherer G, Harley VR. Mutations of the SRY-responsive enhancer of SOX9 are uncommon in XY gonadal dysgenesis. Sex Dev 2010; 4: 321-325
18 Rey RA, Belville C, Nihoul-Fékété C, Michel-Calemard L, Forest MG, Lahlou N, Jaubert F, Mowszowicz I, David M, Saka N, Bouvattier C, Bertrand AM, Lecointre C, Soskin S, Cabrol S, Crosnier H, Léger J, Lortat-Jacob S, Nicolino M, Rabl W, Toledo SP, Baş F, Gompel A, Czernichow P, Josso N. Evaluation of gonadal function in 107 intersex patients by means of serum antimüllerian hormone measurement. J Clin Endocrinol Metab 1999; 84: 627-631
19 Warr N, Greenfield A. The molecular and cellular basis of gonadal sex reversal in mice and humans. Wiley Interdiscip Rev Dev Biol 2012; 1: 559-577
20 Barbaro M, Balsamo A, Anderlid BM, Myhre AG, Gennari M, Nicoletti A, Pittalis MC, Oscarson M, Wedell A. Characterization of deletions at 9p affecting the candidate regions for sex reversal and deletion 9p syndrome by MLPA. Eur J Hum Genet 2009; 17: 1439-1447
21 Ledig S, Hiort O, Wünsch L, Wieacker P. Partial deletion of DMRT1 causes 46,XY ovotesticular disorder of sexual development. Eur J Endocrinol 2012; 167: 119-124
22 Quinonez SC, Park JM, Rabah R, Owens KM, Yashar BM, Glover TW, Keegan CE. 9p partial monosomy and disorders of sex development: review and postulation of a pathogenetic mechanism. Am J Med Genet 2013; 161A: 1882-1896
23 Kim S, Bardwell VJ, Zarkower D. Cell type-autonomous and non-autonomous requirements for Dmrt1 in postnatal testis differentiation. Dev Biol 2007; 307: 314-327
24 Murphy MW, Lee JK, Rojo S, Gearhart MD, Kurahashi K, Banerjee S, Loeuille GA, Bashamboo A, McElreavey K, Zarkower D, Aihara H, Bardwell VJ. An ancient protein-DNA interaction underlying metazoan sex determination. Nat Struct Mol Biol 2015; 22: 442-451
25 Angov E. Codon usage: Nature’s roadmap to expression and folding of proteins. Biotechnol J 2011; 6: 650-659
26 Lin L, Philibert P, Ferraz-de-Souza B, Kelberman D, Homfray T, Albanese A, Molini V, Sebire NJ, Einaudi S, Conway GS, Hughes IA, Jameson JL, Sultan C, Dattani MT, Achermann JC. Heterozygous missense mutations in steroidogenic factor 1 (SF1/Ad4BP, NR5A1 ) are associated with 46,XY disorders of sex sevelopment with normal adrenal function. J Clin Endocrinol Metab 2007; 92: 991-999
27 Tantawy S, Mazen I, Soliman H, Anwar G, Atef A, El-Gammal M, El-Kotoury A, Mekkawy M, Torky A, Rudolf A, Schrumpf P, Grüters A, Krude H, Dumargne M-C, Astudillo R, Bashamboo A, Biebermann H, Köhler B. Analysis of the gene coding for steroidogenic factor 1 (SF1, NR5A1) in a cohort of 50 Egyptian patients with 46,XY disorders of sex development. Eur J Endocrinol 2014; 170: 759-767
28 WuQiang F, Yanase T, Wei L, Oba K, Nomura M, Okabe T, Goto K, Nawata H. Functional characterization of a new human Ad4BP/SF-1 variation, G146A. Biochem Biophys Res Commun 2003; 311: 987-994
29 Wada Y, Okada M, Fukami M, Sasagawa I, Ogata T. Association of cryptorchidism with Gly146Ala polymorphism in the gene for steroidogenic factor-1. Fertil Steril 2006; 85: 787-790
30 Wada Y, Okada M, Hasegawa T, Ogata T. Association of severe micropenis with Gly146Ala polymorphism in the gene for steroidogenic factor-1. Endocr J 2005; 54: 445-448
31 Chen L, Ding XP, Wie X, Li LX. Investigation of mutations in the SRY, SOX9, and DAX1 genes in sex reversal patients from the Sichuan region of China. Genet Mol Res 2014; 13: 1518-1526
32 Barseghyan H, Délot E, Vilain E. New genomic technologies: an aid for diagnosis of disorders of sex development. Horm Metab Res 2015; 47: 312-320