The Genome-wide Association Study Of Chronic Prostatitis/Chronic Pelvic Pain Syndrome And The Study Of The Effect Of Susceptibility Gene LanCL1

Objective:To investigate the role of host genetic factors in the pathogenesis of chronic prostatitis/chronic pelvic pain syndrome（CP/CPPS）,we perform a meta-analysis of two genome-wide association study（GWAS）of CP/CPPS.The effect of significant gene LanCL1,found in the GWAS meta-analysis,in the development of CP/CPPS was explored by using the LanCL1 gene knockout mice.The performance of these studies will reveal the potential complex mechanisms of CP/CPPS and provide effective theoretical foundations for novel target of personalized treatment.Ⅰ.The Genome-Wide Association Study of Chronic Prostatitis/Chronic Pelvic Pain Syndrome.Methods:The GWAS meta-analysis consisted of total 3298 Chinese men aged 20-69 years old from two groups of GWAS.In group 1,GWAS was performed in 1560 men（181 CP/CPPS cases and 1379 controls）from the Fangchenggang area of Guangxi.Group 2 consisted of 1738 men（1513 CP/CPPS cases and 225 controls）from Nanning,Guigang,Qin Zhou and Yulin of Guangxi province.Presence of CP/CPPS was defined by pain score（≥4）of National Institutes of Health Chronic Prostatitis Symptom Index（NIH-CPSI）.Logistics regression model was used in the genome-wide association analyses.The meta-analysis was carried out by PLINK,in which two groups of GWAS result files can be combined in fixed-effects and random-effects result.The enrichment analysis of the differential genes in KEGG,GO and Dis Ge NET database was performed.Results:We identified that the upstream region of LanCL1 exist the most clustering significant SNPs among the SNPs with P-value<1×10^-4.Moreover,there were 3 SNPs（rs4633105,rs6476690,rs12867607）among the significant SNPs have the consistent direction of effect across group 1,group 2 and meta-analysis.The most significant locus associated with CP/CPPS was found on chromosome 8 with the lead SNP rs4633105 in the region of KBTBD11 gene(P=8.219×10^-7).Moreover,rs6476690 and rs12867607 located in the upstream region of ALDH1B1 gene and desert region respectively.Enrichment analysis of differential genes showed that differential genes were mostly enriched in pathways related to nervous system and immune system.Conclusion:KBTBD11,ALDH1B1 and LanCL1 are novel candidate gene that may influence the pathogenesis of CP/CPPS.Genetic factors may influence symptoms of CP/CPPS through affecting the nervous and immune system.Ⅱ.The Study of The Effect of Susceptibility Gene LanCL1 in Gut Microbiota and Central Nervous SystemMethods:We obtained LanCL1 knockout mice using CRISPR/Cas9technology.Wild-type and LanCL1 knockout mice on a normal chow diet were evaluated at 4 weeks and 8–9 weeks of age.16s r RNA sequence and untargeted metabolomics analyses were performed to investigate changes of the gut microbiota and faeces metabolites.Antibiotics treatment was given to both types of mice from 5 to 11 weeks of age in order to delete the influence of gut microbiotas.RT-q PCR,HE staining,AB-PAS staining and the TUNEL assay were performed to detect the alterations in prostate,gut and brain of knockout mice.Results:There was no significant pathological alteration in the prostate of LanCL1 knockout mice.However,the compositions of the gut microbiota and faeces metabolites markedly changed in 8-week-old knockout mice but not in 4-week-old mice.Linear discriminant analysis and t-tests identified Akkermansia as a specific abundant bacteria in knockout mice.N-acetylsphingosine was the most significant downregulated faeces metabolite,which may cause the postponement of neuronal apoptosis.After antibiotics treatment,a significant increase of oxidative damage in the brain of knockout mice was observed,which may have been alleviated by the gut microbiota before.Conclusion:Alterations of the gut microbiota and faeces metabolites alleviated oxidative damage to the brain of LanCL1 knockout mice.Those results suggested that LanCL1,as a genetic factor,may play an important role in the disorders of nervous and mental systems of CP/CPPS,and the microbiota-gut-brain axis can maintain nervous systemic homeostasis in some degree.It also suggested that gut microbiota can be considered as a potential treatment target of CP/CPPS.