Joint utilization of genetic analysis and semi-cloning technology reveals a digenic etiology of Müllerian anomalies

摘要

Identification of a digenic variant combination in MA through genetic analysis and semi-cloning technology. a Deleterious deletion CNVs identified in subjects with MA by CGH microarrays. A novel 1.62 Mb deletion of the human chromosome region 2p24.2 involving GEN1 was identified in subject M45. Recurrent 16p11.2 and 17q12 deletions were identified in five and three subjects with MA, respectively. The genomic deletion regions were covered with green shadow. b The working hypothesis of digenic/oligogenic inheritance modes for MA. The deletion CNVs of GEN1/2p24.2, TBX6/16p11.2 or LHX1/17q12, demonstrate low penetrance in MA, suggesting that a monogenic or Mendelian inheritance mode cannot completely account for the etiology of MA. Apart from single novel or previously identified CNV, another or other genetic variants may simultaneously contribute to MA, thus digenic/oligogenic combinations of variants may reflect the genetic complexity of MA risks. c An additional frameshift mutation (c.6dupC; p.R2fs*55) of WNT9B was identified in subject M45, who carried a GEN1/2p24.2 deletion. These double mutation hits suggest a potential digenic model for the MA in subject M45. d Experimental procedures for producing mouse models of double heterozygous mutants through semi-cloning technology. Wild type AG-haESCs were also injected into oocytes to obtain wild type mice, serving as controls for the mutant founders. e Representative mouse uteri dissected from wild type (WT) and Gen1+/−Wnt9b+/− female mice generated via the semi-cloning technology. Scale bar: 2.5 mm. Mouse uterine lengths (f) or diameters (g) of WT and Gen1+/−Wnt9b+/− female mice generated via the semi-cloning technology. Mouse uterine lengths (h) or diameters (i) of WT, Gen1+/−, Wnt9b+/−, and Gen1+/−Wnt9b+/− female mice generated by the conventional mating way. Female mice in proestrus were collected for analyses. Each side of the uteri was measured and counted. For measurements of diameters, diameter at the middle of each uterus side was measured. All the data are represented as mean ± SEM. One-way ANOVA test, **P < 0.01; ***P < 0.001; n.s. not significant