Novel compound variants of the AR and MAP3K1 genes are related to the clinical heterogeneity of androgen insensitivity syndrome

Abstract

Androgen insensitivity syndrome (AIS; OMIM 300068) is the most frequent cause of 46, XY disorders of sex development (DSD). However, the correlation between genotype and phenotype has not been determined. We conducted a systematic analysis of the clinical characteristics, hormone levels, ultrasonography data and histopathology of a 46, XY Chinese patient with AIS. The family was followed up for nearly 8 years. We applied whole-exome sequencing (WES) for genetic analysis of the pedigree and performed bioinformatic analysis of the identified variants. Human embryonic kidney 293T/17 (HEK293T/17) cells were transiently transfected with wild-type or mutant AR and MAP3K1 plasmid. Cell lysates were used to analyze androgen receptor (AR) production. A novel hemizygous AR variant (c.2070C>A, p. His690Glu) and a rare heterozygous MAP3K1 variant (c.778C>T, p. Arg260Cys) were identified by WES in the proband and her mother. Bioinformatic analysis predicted these two variants to be pathogenic. Multiple amino acid sequence alignments showed that p. His690 and p. Arg260 are conserved among various species. His690Glu is a mutation that decreased the AR production, whereas the Arg260Cys mutation increased the AR production. The novel compound variants of the AR and MAP3K1 genes also increased the production of AR protein. Thus, the phenotype of the patient may be caused by defects in both the AR and MAP3K1 signaling pathways. Compound variants of the AR and MAP3K1 genes resulted in a specific phenotype in this patient with AIS. WES might reveal genetic variants that explain the heterogeneity of AIS.

Nonsense-associated altered splicing of MAP3K1 in two siblings with 46,XY disorders of sex development

“…In summary, this study provides a novel example of NAS, in which a highly potent splice donor site was created in an exon. Furthermore, our data, in conjunction with the previous data indicating the association between MAP3K1 and AR signaling 27 – 29 , imply that the combination of testicular dysgenesis and androgen insensitivity may be a unique phenotype of patients with MAP3K1 abnormalities.…”

“…Therefore, although there is no doubt that testicular dysgenesis is the major cause of 46,XY DSD in patients with MAP3K1 pathogenic variants 7 – 11 , other steps of male sex development may also be affected in these patients. In particular, MAP3K1 abnormalities may perturb the AR signaling in the developing genitalia, because previous studies documented signal crosstalk between MAP3K1 and AR in prostate cancer 27 , 28 and altered AR expression of cells transfected with a mutant MAP3K1 29 . Consistent with this, DHT-induced APOD transactivation, a biological marker of AR function 17 , was less significant in cultured genital skin fibroblasts of our patients compared with that in cells of the control individuals.…”

Genotype-Phenotype Relationship and Follow-up Analysis of a Chinese Cohort With Osteogenesis Imperfecta

“…Using Primer3, version 1.1.4 (http://www.sourceforge.net), and GeneDistiller 2014 (http://www.genedistiller.org/), tagged sequencing primers of the pathogenic genes were designed. The reaction system and the conditions for polymerase chain reaction have been described in a study by Cheng et al 19 Amplicons were sequenced using the ABI 3730 system (Applied Biosystems), and sequence analysis was performed using the autoassembler software Chromas 2.6.6 (Technelysium Pvt Ltd; available at www. technelysium.com.au/chromas.html) and visual inspection.…”

Characteristics and possible mechanisms of 46, XY differences in sex development caused by novel compound variants in NR5A1 and MAP3K1

“…In addition, MAP3K1 is expected to regulate the expression of AR gene [ 19 , 31 , 33 ], and whether MAP3K1 and SRD5A2 (steroid 5 reductase 2, OMIM 607306) [ 41 , 42 ] interact through AR is worthy of further study.…”

“…Using Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific), HEK293T/17 cells were transiently transfected with plasmids containing the wild type NR5A1 (pcDNA3.1- NR5A1 -WT) and MAP3K1 (pcDNA3.1- MAP3K1 -WT) or pcDNA3.1- NR5A1 -MU and pcDNA3.1- MAP3K1 -MU. As previously described [ 19 ], the transfection concentration of a single plasmid was 2.5 ug/mL and the concentration of each plasmid is 1.5 ug/mL at the cotransfection of the two plasmids.…”