Correlation Between CD2AP,TRPC6 Gene Polymorphism And Primary Nephrotic Syndrome In Children Of Zhuang Nationality In Guangxi

Objective:To investigate the correlation between the polymorphism of rs9473132,rs1014001,rs2275446,rs9395287,rs10948368 of CD2AP gene and rs12366144 of TRPC6 gene and the incidence of PNS(primary nephrotic syndrome,PNS)in children of Zhuang nationality in Guangxi;To study the association between the polymorphisms of the above loci and clinical biochemical indexes and serum levels of CD2AP protein.Methods:A case-control study was conducted.A total of 155 newly treated Guangxi Zhuang children with PNS who were admitted to Affiliated Hospital of Youjiang Medical University for Nationalities from July 2018 to September 2020 were selected as the PNS group,and a total of 100 Guangxi Zhuang children with physical examination during the same period were selected as the healthy group.The peripheral blood DNA was extracted,and SNP(single nucleotide polymorphism,SNP)at the intron of CD2AP gene and exon 6 of TRPC6 gene were detected by multiple PCR(polymerase chain reaction,PCR)and second-generation sequencing technology;Serum total protein,albumin,blood urea nitrogen,creatinine,total cholesterol,triglyceride and 24-hour urinary protein quantity were detected by automatic biochemical analyzer;Serum concentrations of CD2AP were detected by ELISA(enzyme-linked immunosorbent assay,ELISA).Statistical analysis was performed using SPSS 26.0 software.The Hardy-Weinberg equilibrium test was used to detect the population representation of the healthy group;Chi-square test was used to analyze the differences in genotype and allele frequency distribution of each SNP locus between the PNS group and the healthy group,and logistic regression analysis was used to predict the risk degree of PNS caused by susceptible SNP sites;T test and Mann-Whitney U test were used to compare the differences in clinical biochemical data and CD2AP protein serum concentration between the PNS group and the healthy group,and ANOVA and Kruskal-Wallis test were used to compare the differences in clinical biochemical data and CD2AP protein serum concentration among the genotypes of the susceptible SNP sites in the PNS group.Results:(1)Eleven loci were found in the introns of CD2AP gene,of which nine loci have been reported by the 1000 Genome Project and two newly discovered loci were located at 47501614 and 47563778 on chromosome 6,respectively.Only rs9473132,rs1014001,rs2275446,rs9395287 and rs10948368 had mutation frequency greater than 1%,which were all present in 255 children in the PNS group and healthy group.One locus was found in exon 6 of TRPC6 gene,namely rs12366144,with mutation frequency greater than 1%,which was found in 255 children of PNS group and healthy group.(2)There were significant differences in the genotype frequencies of rs9473132,rs2275446 and rs9395287 of CD2AP gene between the PNS group and the healthy group(x2=8.344,P=0.015;x2=7.821,P=0.020;x2=6.660,P=0.036).GA genotype at rs9473132,TC genotype at rs2275446,and GA genotype at rs9395287 were negatively correlated with the risk of PNS(OR=0.525,95%CI:0.308-0.898,P=0.019;OR=0.542,95%CI:0.317-0.925,P=0.025;OR=0.552,95%CI:0.323-0.944,P=0.030).(3)There was no significant difEerence in the genotype,dominant model and allele frequency of rs1014001 and rs10948368 of CD2AP gene and rs12366144 of TRPC6 gene between the PNS group and the healthy group.(4)There was no significant difference in serum TP(total protein,TP),ALB(albumin,ALB),BUN(blood urea nitrogen,BUN),CREA(creatinine,CREA),TC(total cholesterol,TC),TG(triglyceride,TG)and 24h Upro(24-hour urinary protein quantity,24h Upro)among the genotypes of CD2AP gene rs9473132,rs2275446 and rs9395287 in the PNS group.(5)There was a statistically significant difference in CD2AP serum concentration between the PNS group and the healthy group(Z=-5.592,P<0.001),but there was no statistically significant difference in CD2AP serum concentration between the rs9473132,rs2275446 and rs9395287 genotypes in the PNS group.(6)There was a strong LD(linkage disequilibrium,LD)among rs9473132,rs1014001,rs2275446,rs9395287 and rs10948368 of CD2AP genes,that is,D’=1,r2>0.33.The above 5 loci constituted three haplotypes:GCTGT,ACCAT and AACAC,and there was no statistical significance between the three haplotypes and the risk of PNS.Conclusion:(1)SNPs of rs9473132,rs2275446 and rs9395287 of CD2AP gene may be related to the incidence of PNS in children of Zhuang nationality in Guangxi,and the GA genotype of rs9473132,TC genotype of rs2275446 and GA genotype of rs9395287 may be protective genotypes to reduce the incidence of PNS.(2)SNPs of rs1014001 and rs109483 68 of CD2AP gene and rs12366144 of TRPC6 gene may not be associated with the incidence of PNS in children of Zhuang nationality in Guangxi.(3)SNPs of rs9473132,rs2275446 and rs9395287 of CD2AP gene may not affect serum levels of TP,ALB,BUN,CREA,TC,TG and 24h Upro in Guangxi Zhuang children with PNS.(4)CD2AP protein may be a renal protective protein,but its expression level may not be regulated by rs9473132,rs2275446 and rs9395287.(5)There was strong LD among rs9473132,rs1014001,rs2275446,rs9395287 and rs10948368 of CD2AP gene,but their haplotypes may not affect the incidence of PNS in children of Guangxi Zhuang nationality.