| BackgroundIn recent years,the incidence of diabetes has continued to increase,and its common complication,diabetic nephropathy(DN),has a prevalence of 20% to 40%in patients with type 2 diabetes mellitus(T2DM),which causes a heavy economic burden for society.However,the pathogenesis of DN is complex and has not yet been fully elucidated.Type 2 DN is a complex disease caused by the interaction between gene effects and environmental factors.Epigenetic mechanisms play an important role in its development.However,DN epigenetics research is still in its infancy,and there is no research on the methylation status of renal tissue in patients with DN.Therefore,this study intends to use the kidney tissue of DN patients to map DNA methylation,and combined with bioinformatics analysis to explore the specific methylation changes and related signaling pathways that occur during DN development,and lay a foundation for the exploration of therapeutic targets.Purpose1.Draw the genome-wide DNA methylation map of kidney tissue in patients with DN for the first time,and provide evidence for subsequent research on epigenetics of DN patients;2.Through the drawing of DNA methylation map and bioinformatics analysis,screen the unique methylation difference sites in several type 2 DN patients,which is helpful for the exploration of DN therapeutic targets,thus laying the foundation for precise treatment and individualized non-invasive diagnosis.Methods1.Source of specimen: 7 non-diabetic patients(paracancerous tissues),7 T2 DM patients(paracancerous tissues),7 patients with type 2 DN(kidney puncture tissue)and 8 patients with type 2 non-diabetic renal disease(NDRD)(including 3 cases of membranous nephropathy,3 cases of Ig A nephropathy and 2 cases of focal stage glomerulosclerosis,all of which were renal puncture tissue),the diagnostic criteria were confirmed by renal pathological examination;2.Whole-genome methylation sequencing and methylation mapping: completed whole-genome methylation sequencing and DNA methylation mapping with Illumina's second-generation high-throughput sequencing platform combined with whole-genome bisulfite treatment;3.Bioinformatics analysis: through methylation difference analysis and GO and KEGG analysis,pathways closely related to DN were obtained,and several genes related to the pathogenesis of DN were anchored.Results1.We drew the genome-wide DNA methylation map of kidney tissue in patients with DN for the first time;2.649 methylation difference regions between DN group and NDRD group were found and 609 methylation difference regions were found between DN group and DM group.After comprehensive analysis,there were 160 methylation difference regions in which the DN group different from both the NDRD group and the DM group;3.Combined with methylation level difference analysis and related enrichment analysis,it was found that hippo signaling pathway,pantothenate,Co A biosynthesis,endocytosis and cell adhesion molecules and other biochemical metabolic pathways and signal transduction pathway were most correlated with DN group;4.Combined with the degree of methylation difference and enrichment,we further anchored the 7 most ranked genes associated with the pathogenesis of DN:CTGF,MMP-9,MTHFR,let-7a-3,ITGB1,PANK1 and Mst1.ConclusionHippo signaling pathway,pantothenate,Co A biosynthesis,endocytosis and cell adhesion molecules play an important role in the development of DN.CTGF,MMP-9,MTHFR,let-7a-3,ITGB1,PANK1 and Mst1 genes rank highest in the degree of methylation difference and enrichment in the DN group,which may become the therapeutic targets or non-invasive diagnostic markers of DN in the future. |
PDF Full Text Request