Spatiotemporal dynamics of androgen signaling underlie sexual differentiation and congenital malformations of the urethra and vagina

Disorders of sex development (DSDs) are congenital anomalies that affect sexual differentiation of genitourinary organs and secondary sex characters. A common cause of female genital virilization is congenital adrenal hyperplasia (CAH), in which excess androgen production during development of 46XX females can result in vaginal atresia, masculinization of the urethra, a single urogenital sinus, and clitoral hypertrophy or ambiguous external genitalia. Development of the vagina depends on sexual differentiation of the urogenital sinus ridge, an epithelial thickening that forms where the sex ducts attach to the anterior urethra. In females, the sinus ridge descends posteriorly to allow the vaginal opening to form in the vulva, whereas in males and in females with CAH, androgens inhibit descent of the sinus ridge. The mechanisms that regulate development of the female urethra and vagina are largely unknown. Here we show that the timing and duration of, and the cell population targeted by, androgen signaling determine the position of vaginal attachment to the urethra. Manipulations of androgen signaling in utero reveal a temporal window of development when sinus ridge fate is determined. Cell type-specific genetic deletions of androgen receptor (Ar) identify a subpopulation of mesenchymal cells that regulate sinus ridge morphogenesis. These results reveal a common mechanism that coordinates development of the vagina and feminization of the urethra, which may account for development of a single urogenital sinus in females exposed to excessive androgen during a critical period of prenatal development.

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Characterising SRD5A2 Gene Variants in 37 Indonesian Patients with 5-Alpha-Reductase Type 2 Deficiency

International Journal of Endocrinology ◽

10.1155/2019/7676341 ◽

2019 ◽

Vol 2019 ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

Nanis S. Marzuki ◽

Firman P. Idris ◽

Hannie D. Kartapradja ◽

Alida R. Harahap ◽

Jose R. L. Batubara

Keyword(s):

Autosomal Recessive ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Compound Heterozygous ◽

Phenotypic Characteristics ◽

Sex Development ◽

Srd5a2 Gene ◽

Diagnostic Approaches ◽

Autosomal Recessive Condition

The 5-alpha-reductase type 2 deficiency (5ARD2) is an autosomal recessive condition associated with impairment in the conversion of testosterone to dihydrotestosterone. This condition leads to undervirilisation in 46,XY individuals. To date, there have been more than 100 variations identified in the gene responsible for 5ARD2 development (steroid 5-alpha-reductase 2, SRD5A2). However, few studies have examined the molecular characterisation of Indonesian 5ARD2 cases. In the current study, we analysed 37 subjects diagnosed with 46,XY DSD (disorders of sex development) with confirmed variations in the SRD5A2 gene. We examined results from testosterone/dihydrotestosterone (T/DHT) and urinary etiocholanolone/androsterone (Et/An) ratios, as well as from molecular and clinical analyses. Twelve variants in the SRD5A2 gene were identified, and 6 of which were novel, namely, c.34–38delGinsCCAGC, p.Arg50His, p.Tyr136∗, p.Gly191Arg, p.Phe194Ile, and p.Ile253Val variants. Moreover, we determined that 20 individuals contained harmful mutations, while the remaining 17 variants were benign. Those containing harmful mutations exhibited more severe phenotypes with median external genitalia masculinisation scores (EMS) of 3 (1.5–9) and were more likely to be diagnosed at a later age, reared as female, and virilised at pubertal age. In addition, the respective sensitivities for detecting severe 5ARD2 cases using T/DHT (cutoff: 10) and urinary Et/An ratios (cutoff: 0.95) were 85% and 90%, whereas mild cases were only identified with 64.7% and 47.1% sensitivity, respectively. Although we were unable to identify clear correlations between genotypic and phenotypic characteristics in this study, we clearly showed that individuals who were homozygous or compound heterozygous for any of the harmful mutations were more likely to exhibit classic 5ARD2 phenotypes, lower EMS, female assignment at birth, and virilisation during puberty. These results serve to inform the development of improved clinical and molecular 5ARD2 diagnostic approaches, specifically in Indonesian patients.

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Comprehensive Transcriptome Analysis of Hypothalamus Reveals Genes Associated With Disorders of Sex Development in Pigs

10.21203/rs.3.rs-45383/v1 ◽

2020 ◽

Author(s):

Shuwen Tan ◽

Yi Zhou ◽

Haiquan Zhao ◽

Jinhua Wu ◽

Hui Yu ◽

...

Keyword(s):

Expression Profiles ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Rna Isolation ◽

Functional Enrichment ◽

Regulation Mechanism ◽

Normal Female ◽

Sex Development ◽

Mirna Expression Profiles ◽

Hormone Biosynthesis

Abstract Background Disorders of sex development (DSD) is a chronic autoimmune disease characterized by systemic inflammation, pathological osteogenesis, and endocrine abnormality. However, its genetic etiology remains mostly unknown. In addition, little research focuses on the regulation mechanism from the view of transcriptomics in the hypothalamic-pituitary-gonadal axis (HPGA). The hypothalamus is the integrated center of the HPGA mediating neural, hormonal, and environmental stimulus to sex development. Methods Three XX-DSD (SRY-negative) pig (DSD) and three NF pigs (five months old, 40 kg ± 5 kg) were selected by external genitalia observation and sex determining region Y gene (SRY) detection. The hypothalamus were sampled for RNA isolation, and the mRNA, lncRNA and miRNA expression profiles were analyzed by sequencing. Results A total of 1,258 lncRNAs, 1,086 mRNAs, and 61 microRNAs were found to differentially express in XX-DSD pigs compared with normal female pigs. Many genes in hormone biosynthesis and secretion pathway are significantly up-regulated, and the up-regulation of GNRH1, KISS1 and AVP may be the candidate genes leading the abnormal secretion of GnRH. Next, we predicted the lncRNA-miRNA-mRNA co-expression triplets and constructed three competing endogenous RNA (ceRNA) potentially associated with DSD. Functional enrichment suggested TCONS_00340886, TCONS_00000204 and miR-181a were related to GnRH secretion. Conclusions Our research revealed the first transcriptomic profile in the hypothalamus of XX-DSD pigs and provided new insight in coding and non-coding RNAs that may be associated with DSD in pigs.

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MAMLD1 and Differences/Disorders of Sex Development: An Update

Sexual Development ◽

10.1159/000519298 ◽

2021 ◽

pp. 1-12

Author(s):

Mami Miyado ◽

Maki Fukami ◽

Tsutomu Ogata

Keyword(s):

Leydig Cell ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Gene Interactions ◽

Testicular Function ◽

Ovarian Dysfunction ◽

Causative Gene ◽

Fetal Testis ◽

Sex Development ◽

Age Dependent

MAMLD1 (alias CXorf6) was first documented in 2006 as a causative gene of 46,XY differences/disorders of sex development (DSD). MAMLD1/Mamld1 is expressed in the fetal testis and is predicted to enhance the expression of several Leydig cell-specific genes. To date, hemizygous MAMLD1 variants have been identified in multiple 46,XY individuals with hypomasculinized external genitalia. Pathogenic MAMLD1 variants are likely to cause genital abnormalities at birth and are possibly associated with age-dependent deterioration of testicular function. In addition, some MAMLD1 variants have been identified in 46,XX individuals with ovarian dysfunction. However, recent studies have raised the possibility that MAMLD1 variants cause 46,XY DSD and ovarian dysfunction as oligogenic disorders. Unsolved issues regarding MAMLD1 include the association between MAMLD1 variants and 46,XX testicular DSD, gene-gene interactions in the development of MAMLD1-mediated DSD, and intracellular functions of MAMLD1.

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AMH and AMHR2 Involvement in Congenital Disorders of Sex Development

Sexual Development ◽

10.1159/000518273 ◽

2021 ◽

pp. 1-9

Author(s):

Franco G. Brunello ◽

Rodolfo A. Rey

Keyword(s):

Molecular Genetic ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Fetal Life ◽

Fallopian Tubes ◽

Female Fetus ◽

Sequencing Technologies ◽

Sex Development ◽

Mullerian Ducts ◽

Gene Maps

Anti-müllerian hormone (AMH) is 1 of the 2 testicular hormones involved in male development of the genitalia during fetal life. When the testes differentiate, AMH is secreted by Sertoli cells and binds to its specific receptor type II (AMHR2) on the müllerian ducts, inducing their regression. In the female fetus, the lack of AMH allows the müllerian ducts to form the fallopian tubes, the uterus, and the upper part of the vagina. The human AMH gene maps to 19p13.3 and consists of 5 exons and 4 introns spanning 2,764 bp. The AMHR2 gene maps to 12q13.13, consists of 11 exons, and is 7,817 bp long. Defects in the AMH pathway are the underlying etiology of a subgroup of disorders of sex development (DSD) in 46,XY patients. The condition is known as the persistent müllerian duct syndrome (PMDS), characterized by the existence of a uterus and fallopian tubes in a boy with normally virilized external genitalia. Approximately 200 cases of patients with PMDS have been reported to date with clinical, biochemical, and molecular genetic characterization. An updated review is provided in this paper. With highly sensitive techniques, AMH and AMHR2 expression has also been detected in other tissues, and massive sequencing technologies have unveiled variants in AMH and AMHR2 genes in hitherto unsuspected conditions.

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Molecular Characteristics of Sequence Variants in GATA4 in Patients with 46,XY Disorders of Sex Development without Cardiac Defects

Sexual Development ◽

10.1159/000511258 ◽

2019 ◽

Vol 13 (5-6) ◽

pp. 240-245

Author(s):

Jin-Ho Choi ◽

Yena Lee ◽

Arum Oh ◽

Gu-Hwan Kim ◽

Han-Wook Yoo

Keyword(s):

Heart Defects ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Pathogenic Mutation ◽

Luciferase Reporter ◽

Molecular Characteristics ◽

Sequence Variants ◽

Sex Development ◽

Stimulation Tests

A GATA4 haploinsufficiency has been well described in patients with congenital heart defects (CHDs), whilst only a few studies have reported mutations related to a 46,XY disorder of sex development (DSD) phenotype. This study investigated the clinical phenotypes and molecular characteristics of two 46,XY DSD patients harboring GATA4 variants. Mutation analysis was performed using a targeted gene panel or whole-exome sequencing. The transactivation activity of each variant protein was examined by in vitro luciferase reporter assay using the AMH and SRY promoters. Subject 1 presented with a micropenis and hypospadias. Subject 2 showed complete female external genitalia with a 46,XY karyotype. Both patients were responsive to hCG stimulation tests and did not manifest CHD. A novel heterozygous variant, c.643A>G (p.R215G), in GATA4 was identified in Subject 1, whereas Subject 2 harbored a previously reported variant, c.1220C>A (p.P407Q), in GATA4 and a previously known pathogenic mutation, i.e., c.226C>T (p.Q76*) in the AR gene. The reporter assays using the SRY and AMH promoters revealed decreased transcriptional activity of both p.P407Q and p.R215G. However, the GATA4 p.P407Q variant was classified as likely benign. In conclusion, it is essential to integrate clinical features and endocrine findings when interpreting sequence variants.

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46,XX DSD: Developmental, Clinical and Genetic Aspects

Diagnostics ◽

10.3390/diagnostics11081379 ◽

2021 ◽

Vol 11 (8) ◽

pp. 1379

Author(s):

Camelia Alkhzouz ◽

Simona Bucerzan ◽

Maria Miclaus ◽

Andreea-Manuela Mirea ◽

Diana Miclea

Keyword(s):

Phenotypic Variability ◽

Disorders Of Sex Development ◽

Life Quality ◽

Prenatal Development ◽

Abnormal Development ◽

Postnatal Exposure ◽

Sex Development ◽

Genetic Abnormalities ◽

Proper Diagnosis

Differences in sex development (DSD) in patients with 46,XX karyotype occur by foetal or postnatal exposure to an increased amount of androgens. These disorders are usually diagnosed at birth, in newborns with abnormal genitalia, or later, due to postnatal virilization, usually at puberty. Proper diagnosis and therapy are mostly based on the knowledge of normal development and molecular etiopathogenesis of the gonadal and adrenal structures. This review aims to describe the most relevant data that are correlated with the normal and abnormal development of adrenal and gonadal structures in direct correlation with their utility in clinical practice, mainly in patients with 46,XX karyotype. We described the prenatal development of structures together with the main molecules and pathways that are involved in sex development. The second part of the review described the physical, imaging, hormonal and genetic evaluation in a patient with a disorder of sex development, insisting more on patients with 46,XX karyotype. Further, 95% of the etiology in 46,XX patients with disorders of sex development is due to congenital adrenal hyperplasia, by enzyme deficiencies that are involved in the hormonal synthesis pathway. The other cases are explained by genetic abnormalities that are involved in the development of the genital system. The phenotypic variability is very important in 46,XX disorders of sex development and the knowledge of each sign, even the most discreet, which could reveal such disorders, mainly in the neonatal period, could influence the evolution, prognosis and life quality long term.

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Disorders of Sex Development: Can You Be Sure This Baby Is A Boy or Girl? We Must See Beyond The Diagnosis

Journal of Shaheed Suhrawardy Medical College ◽

10.3329/jssmc.v10i2.41170 ◽

2019 ◽

Vol 10 (2) ◽

pp. 103-110

Author(s):

Mohammed Shadrul Alam ◽

Mirza Kamrul Zahid ◽

Paritosh Kumar Palit ◽

Abhi Kumar Chakraborty ◽

Nirupama Saha ◽

...

Keyword(s):

Disorders Of Sex Development ◽

External Genitalia ◽

Sex Development ◽

Newborn Children

Throughout the pregnancy, the parents have anticipated whether their child will be a boy or a girl. No part of a newborn baby’s anatomy arouses as much interest initially as the external genitalia. Most newborn children have the typical features of a boy or girl, but in some cases the baby’s sex can’t be clearly identified. Infants born with ambiguous or abnormal genitalia may have indeterminate phenotypic sex.1 Disorders of sex development (DSDs), formerly termed intersex conditions, are congenital conditions in which development of the chromosomal, gonadal, or anatomic sex is atypical and may affect up to 1:1000 individuals in the population.2 J Shaheed Suhrawardy Med Coll, December 2018, Vol.10(2); 103-110

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Genetic Analysis for the Diagnosis of Disorders of Sexual Development in Indonesia

Journal of Biomedicine and Translational Research ◽

10.14710/jbtr.v2i2.622 ◽

2016 ◽

Vol 2 (2) ◽

pp. 44

Author(s):

Sultana MH Faradz

Keyword(s):

Sexual Development ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Gonadal Development ◽

Androgen Insensitivity Syndrome ◽

Medical Professionals ◽

Adrenal Hyperplasia ◽

Disorders Of Sexual Development ◽

Sex Development

Disorders of sex development (DSD) is defined by congenital conditions in which development of chromosomal, gonadal, or anatomical sex is atypical, while in clinical practice this term means any abnormality of the external genitalia. DSD patients have been managed by a multidisciplinary gender team in our center as collaboration between Dr. Kariadi province referral hospital and Faculty of Medicine Diponegoro University. Diagnosis should be established by specific physical examination hormonal, chromosomal and DNA studies; and imaging for most of the cases depending on indication.Since 2004 the involvement of molecular and cytogenetic analysis so far can diagnosed many of the DSD cases. Most of the genetically proven cases were Congenital Adrenal hyperplasia, Androgen Insensitivity syndrome and sex chromosomal DSD that lead abnormal gonadal development. Many of them remain undiagnosed, further testing such as advanced DNA study should be carried out in collaboration with other center in overseas.The novel genes were found in some cases that contributed for the management of DSD. Information for medical professionals, patients, family members and community about the availability and necessity of DSD diagnosis should be delivered to improve DSD management and patient quality of life.

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Disorders of Sex Development of Adrenal Origin

Frontiers in Endocrinology ◽

10.3389/fendo.2021.770782 ◽

2021 ◽

Vol 12 ◽

Author(s):

Gabriela P. Finkielstain ◽

Ana Vieites ◽

Ignacio Bergadá ◽

Rodolfo A. Rey

Keyword(s):

Congenital Adrenal Hyperplasia ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Special Focus ◽

Adrenal Hyperplasia ◽

Hydroxylase Deficiency ◽

Trophic Effect ◽

Salt Wasting ◽

Sex Development ◽

21 Hydroxylase Deficiency

Disorders of Sex Development (DSD) are anomalies occurring in the process of fetal sexual differentiation that result in a discordance between the chromosomal sex and the sex of the gonads and/or the internal and/or external genitalia. Congenital disorders affecting adrenal function may be associated with DSD in both 46,XX and 46,XY individuals, but the pathogenic mechanisms differ. While in 46,XX cases, the adrenal steroidogenic disorder is responsible for the genital anomalies, in 46,XY patients DSD results from the associated testicular dysfunction. Primary adrenal insufficiency, characterized by a reduction in cortisol secretion and overproduction of ACTH, is the rule. In addition, patients may exhibit aldosterone deficiency leading to salt-wasting crises that may be life-threatening. The trophic effect of ACTH provokes congenital adrenal hyperplasia (CAH). Adrenal steroidogenic defects leading to 46,XX DSD are 21-hydroxylase deficiency, by far the most prevalent, and 11β-hydroxylase deficiency. Lipoid Congenital Adrenal Hyperplasia due to StAR defects, and cytochrome P450scc and P450c17 deficiencies cause DSD in 46,XY newborns. Mutations in SF1 may also result in combined adrenal and testicular failure leading to DSD in 46,XY individuals. Finally, impaired activities of 3βHSD2 or POR may lead to DSD in both 46,XX and 46,XY individuals. The pathophysiology, clinical presentation and management of the above-mentioned disorders are critically reviewed, with a special focus on the latest biomarkers and therapeutic development.

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Disorders of sex development: a genetic study of patients in a multidisciplinary clinic

Endocrine Connections ◽

10.1530/ec-14-0085 ◽

2014 ◽

Vol 3 (4) ◽

pp. 180-192 ◽

Cited By ~ 10

Author(s):

Luigi Laino ◽

Silvia Majore ◽

Nicoletta Preziosi ◽

Barbara Grammatico ◽

Carmelilia De Bernardo ◽

...

Keyword(s):

Sex Chromosome ◽

Genetic Defect ◽

Disorders Of Sex Development ◽

External Genitalia ◽

Molecular Study ◽

Testicular Development ◽

Androgen Synthesis ◽

Sex Development ◽

Dna Alterations ◽

Definition Of

Sex development is a process under genetic control directing both the bi-potential gonads to become either a testis or an ovary, and the consequent differentiation of internal ducts and external genitalia. This complex series of events can be altered by a large number of genetic and non-genetic factors. Disorders of sex development (DSD) are all the medical conditions characterized by an atypical chromosomal, gonadal, or phenotypical sex. Incomplete knowledge of the genetic mechanisms involved in sex development results in a low probability of determining the molecular definition of the genetic defect in many of the patients. In this study, we describe the clinical, cytogenetic, and molecular study of 88 cases with DSD, including 29 patients with 46,XY and disorders in androgen synthesis or action, 18 with 46,XX and disorders in androgen excess, 17 with 46,XY and disorders of gonadal (testicular) development, 11 classified as 46,XX other, eight with 46,XX and disorders of gonadal (ovarian) development, and five with sex chromosome anomalies. In total, we found a genetic variant in 56 out of 88 of them, leading to the clinical classification of every patient, and we outline the different steps required for a coherent genetic testing approach. In conclusion, our results highlight the fact that each category of DSD is related to a large number of different DNA alterations, thus requiring multiple genetic studies to achieve a precise etiological diagnosis for each patient.

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