Risk Of Rotator Cuff Tears And The Polymorphisms And Mechanism Of DEFB1,ESRRB,and FGFs Genes

Background:Increasing studies have indicated a prominent correlation between genetic factors and the risk of rotator cuff tears(RCT)recently.In China,limited investigations have been conducted on polymorphisms of susceptibility genes associated with RCT.The present case-control study aimed to analyze the correlation between gene polymorphisms of DEFB1,estrogen-related receptor b(ESRRB),and fibroblast growth factors(FGFs)and the occurrence of RCT risk in Chinese Han nationality.Methods:A total of 100 RCT patients and 116 healthy persons were recruited and their peripheral blood samples were collected.19 SNP loci of DEFB1,ESRRB,and FGFs genes were genotyped,and genetic association analysis was carried out,including single nucleotide polymorphism(SNP),linkage disequilibrium(LD),and haploid.A model of multivariate logistic regression was subsequently constructed.In order to investigate the regulation and control effect of SNP on tendon regeneration and the molecular mechanism,DEFB1(CC)and DEFB1(GG)vectors were constructed respectively,transfected with rat tendon stem cells.Differential binding proteins were screened for both different genotypes using DNA pull down and mass spectrometry,and the function of the binding proteins was further validated.Results:The results showed that rs4903399 genotype was significantly different between the two groups(CC vs.CT+TT)(OR:0.49;95%CI:0.26-0.89;P=0.02),rs11750845 was associated with increased risk of RCT(CC vs.CT+TT)(OR:0.55;95%CI:0.31-0.96;P=0.04),while RS13317 was associated with reduced risk of RCT CC vs.CT+TT(OR:2.64;95%CI:1.20-6.28;P=0.02).The CT genotype of RS4903399 was significantly different between the two groups(P=0.02),and the dominant model showed that RS4903399 was an independent factor of RCT(CC vs.CT+TT)(OR:0.40;95%CI:0.21-0.77;P=0.01).The dominant model of RS13317 showed a significant trend(CC vs.CT+TT)(OR:2.30;95%CI:0.98-5.41;P=0.05),CTCCTCGA of ESRRB was associated with reduced risk of RCT(OR:0.23;95%CI:0.06-0.86;P=0.02),CTCCTTGA increased the risk of RCT(OR:4.70;95%CI:1.26-17.52;P=0.01).In FGF3,we found that both CAAG(P<0.05)and TGGA(P<0.05)haploid types increased the risk of RCT.Similarly,we found that the haploid CTCT type of FGF10 was significantly associated with the occurrence of RCT(OR:0.64;95%CI:0.43-0.94;P=0.03).In the cell experiment of DEFB1,we found that DEFB1 overexpression could promote the expression of Scx,TNC,COL i,COL iii and other tendinous markers,and inhibit the expression of OPN and Sox9 osteogenic markers.At the same time,DEFB1 GG(wild type)is more easily expressed than CC(variant type).However,its CC(variant)affects DEFB1 expression by weakening its binding ability to FUS target protein.FUS protein can positively regulate the expression of DEFB1,thereby promoting the expression of Scx,TNC,COL i,COL iii and other tendinous markers,and inhibiting the expression of OPN and Sox9 osteogenic markers.When FUS protein expression is down-regulated,cell staining shows significantly increased calcification.Conclusions:The results of this study confirmed the correlation between DEFB1,ESRRB,FGFs and RCT in Chinese Han population,provided valuable evidence for the relationship between rotator cuff tear and gene polymorphism,and clarified the pathogenesis of the effect of DEFB1 gene polymorphism on the risk of rotator cuff tear.However,further large samples and well-designed experiments are needed to determine the correlation.