The Relationships Between DNAH1,DNAH5,DNAH11 Gene Variations And Asthenozoospermia

Objective: From the perspective of molecular genetics,this study aimed to screen out the potential pathogenic variations of DNAH1,DNAH5 and DNAH11 genes among asthenozoospermia patients,based on the targeted high-throughput sequencing technology and bioinformatics analysis,and explore the relationship between these gene variations and asthenozoospermia.The variation sites were explored for constructing the genetic diagnosis platform of primary asthenozoospermia with clinial practical value.Methods: A total of 143 diopathic athenozoospermia patients diagnosed in Reproductive Center and Prenatal Diagnostic Center of the First Hospital of Jilin University were enrolled as main subjects.Inclusion criteria were as follows: no severe varicocele;no previous medical history of infection,genital trauma,major diseases,etc;no previous history of radiation exposure or radiotherapy or chemotherapy;normal chromosome karyotype;no AZF microdeletion;no hyperthermia during the treatment period;Han population in northeast China.In this study,basic information such as age and stertility years were collected by questionnaires.Routine semen analysis,peripheral blood serum reproductive hormone detection,chromosome karyotype analysis and AZF microdeletion detection were conducted among those enrolled subjects.The exonic regions of target genes including DNAH1,DNAH5,and DNAH11 genes were sequenced by targeted high-throughput sequencing technology.All the detected variations were verified by Sanger sequencing.On the basis of online software such as SIFT,Polyphen-2,Mutaiontaster2 and Human splicing Finder 3.1,as well as related databases like db SNP,1000 genomes,Exome variation Server,Exome Aggregation Consortium and etc.,this study further conducted bioinformatics analysis on the variations.Results are as follows:1.Targeted high-throughput sequencing technology was performed on 143 patients,and a total of 5 patients were found to carry pathogenic variations of DNAH1,DNAH5,or DNAH11 genes,accounting for approximate 3.50%?5/143?of the included population.Thereinto,three patients were found to have pathogenic SNV associated with DNAH1 gene,accounting for 2.10%?3/143?;one of 143?0.70%?patients was detected with potentially pathogenic SNV in DNAH5 gene;and 2 patients were identified with DNAH11 gene related pathogenic SNV,about1.40%?2/143?.2.The detected variations in DNAH1 included c.7066C>T?rs780050981?,c.11726₁1727del?rs779490893?and c.8322+3del.Among them,the novel variation c.8322+3del was predicted by HSF3 as Most probably affect splicing,with pathogenicity;the variation c.7066C>T?rs780050981?was predicted to affect the function of DNAH1 protein by SIFT,polyphen-2,and Mutationtaster;variation c.11726₁1727del was exclusively identified as pathogenic by Mutationtaster.3.The detected variations in DNAH5 gene were c.3502G>A?COSM232178?and c.2578-11₂578-7del?rs758852668?,both of which were carried by one patient in this study.Bioinformatics analysis showed that the two variations were weakly pathogenic,in accordance with mild abnormal sperm motility of the patient.4.The detected variations in DNAH11 genes involved c.13171C>T?rs761855200?,c.9484-1g >T,and c.12428T> c?rs751994566?.New variation c.9484-1g >T was assessed by HSF3 to affect splicing of post-transcriptional m RNA of DNAH11 gene;the variation c.13171C>T was a nonsense mutation,predicted by Mutationtaster as Disease causing,and had potential pathogenicity;Variation c.12428T> c was evaluated to be pathogenic by polyphen-2 and Mutationtaster.5.Proved by the existing databases and relevant literature reports,the above 8variation loci have potential pathogenicity.Conclusions:1.The detection rate of DNAH1,DNAH5,DNAH11 gene variations in asthenozoospermia patients was low,which may be consistent with their highly conserved characteristics.2.Two novel variations DNAH1 c.8322+3del and DNAH11 c.9484-1g >T were potentially associated with the occurrence of asthenozoospermia.3.Six other variations DNAH1 c.7066C>T?rs780050981?,DNAH1 c.11726₁1727del?rs779490893?,DNAH5 c.3502G>A?COSM232178?,DNAH5 c.2578-11₂578-7del?rs71600031?,DNAH11 c.13171C>T?rs761855200?,DNAH11 c.12428T>C?rs751994566?may also be associated with asthenozoospermia.