Genotype,Clinical Phenotype And Spermatogenesis In Patients With Congenital Hypogonadotropic Hypogonadism(CHH),Focusing On Gene Mutation Of FGFR1

Objective.The aim of this study was to explore the relationship between genotype,clinical phenotype and effect of sperm-inducing therpay in congenital hypogonadotropic hypogonadism(CHH)patients,analyze the sperm-inducing effect of patients with FGFR1 mutation,and explore the effect of FGFR1 mutation on spermatogonial function.Methods.Next-generation sequencing(NGS)was used to identify the mutation genes carried by CHH patients.Clinical data and laboratory tests were collected to compare the efficacy of sperm-inducing therapy in patients with different mutations,including FGFR1 gene mutation.Transcriptome sequencing,scratch test and CCK-8 proliferation test were used to explore the effect of FGFR1 gene defect on cell function.Results.1)In the first part,158 CHH patients were sequenced.It was found that 53.16%of CHH patients carried CHH related gene mutation.FGFR1(15.19%),PROKR2(10.13%)and ANOS 1(6.96%)were the most common mutated genes in CHH patients.The incidence of cryptorchidism in patients with FGFR1 mutation was 43.75%.CHH patients with gene mutation,especially those with P/LP mutation,have smaller baseline testicular volume and longer median time for first sperm appearance.2)In the second part,14 patients with FGFR1 gene mutation and 25 patients without gene mutation were included.The incidence of cryptorchidism was 50.00%(7/14)and 12.00%(3/25),respectively(P=0.019).The baseline testicular volume was 1.60(0.50-2.00)ml and 2.00(1.75-4.00)ml,respectively(P=0.033).The cumulative success rate of spermatogenesis in the two groups was 35.71%and 68.75%respectively(P=0.047).The median time of spermatogenesis in FGFR1 mutation group was longer than that in mutation negative group(16 vs.10 months).3)In the third part,FGFR1 knockdown was carried out in Mouse Spermatogonia by shRNA packaged with lentivirus.RT-qPCR and Western blot were used to verify the knockdown efficiency.Transcriptome sequencing analysis revealed 1522 different gene between the knock down group and empty vector group.The migration and proliferation ability of GC-1 spg cell line with FGFR1 knockdown were weakend than those of empty vector group.Conclusion.Patients with mutation,especially those with P/LP gene mutation,had more severe reproductive phenotype damage and need longer time for first sperm appearance.The patients with FGFR1 gene mutation had a higher prevalence of cryptorchidism and smaller testicular volume.The patients with FGFR1 gene mutation need longer treatment for first sperm apperance,and the cumulative success rate of spermatogenesis in one year was lower than that in the patients without gene mutation.FGFR1 gene mutation leads to dysfunction of spermatogonia and changes in gene expression pattern,which provides a remedial target for further treatment of testicular injury caused by FGFR1 gene mutation.