| This thesis uses single-cell sequencing technology to map the gene expression profiles of each subgroup of blood T cells from 8 patients with Parkinson’s disease and 6 normal controls,and integrates genotype data to study the immune regulation mechanism of Parkinson’s disease-related gene expression.The immunotherapy of Parkinson’s disease provides potential targets.This experiment uses whole-genome sequencing technology to genotype patients and normal people,and uses PLINK1.9 software to control the quality of genomic mutations,and further combines with existing literature studies to screen out strong linkage with Parkinson’s susceptible mutations.Variations in heritabi lity,a total of 4,348 single nucleotide polymorphisms(SNPs)were used for subsequent gene expression quantitative trait loci(eQTL)research.This experiment uses the R language Seurat function to perform dimensionality reduction,clustering,and cell type annotation on the single cells of T immune cells in all samples.According to the measured gene expression profile and the characteristic expression genes of each subgroup of T cells,the T cells are Annotated as 5 major subtypes(including 22 more detailed subgroups),the 5 major subtypes are:CD4~+T cells,CD8~+T cells,mucosal-associated invariant T cells(MAIT),γ-δT cells(gd T)and double negative T cells(DN).This experiment uses the Metrix EQTL function of R research to perform single-cell gene expression regulation analysis to identify T cell type-specific and disease-specific eQTLs related to Parkinson’s disease.In the cell type specific analysis process,31,339,336,600 and 474 cell type specific gene expression regulation were obtained in CD4~+T cells,CD8~+T cells,MAI T cells,gd T cells and DN T cells,respectively The rs11950533 mutation in CD4~+T cells regulates the CAMLG gene,the rs11577197 mutation in CD8~+T cells regulates the VAMP4 gene,and the rs8087969 mutation in MAI T cells regulates the expression of the MEX3C gene as examples to illustrate that Parkinson’s disease susceptibility mutations may be regulated by specific The gene expression of different types of immune cells affects the occurrence and development of the disease;in the disease-specific eQTL analysis,by comparing the difference in gene expression regulation between normal people and patients with Parkinson’s disease,screening for Parkinson’s disease-specific eQTL Point,a total of 10,015 disease-specific gene expression regulation relationships.Among them,CD4~+T cells,CD8~+T cells,MAI T cells,gd T cells,and DN T cells detected 726,1164,3729,2766,5321 on the regulation of gene expression,through the relationship with Parkinson’s disease susceptibility genes and Susceptible SNPs were compared.Four of the regulatory relationships were analyzed in this experiment:rs3104784 regulates TUFM gene,rs2271613regulates PGS1 gene,rs9271222 regulates HLA-DRB5 gene and rs4690198regulates GAK gene.In order to further clarify the expression regulation mechanism of Parkinson’s susceptibility genes,this study used disease-specific eQTL regulatory genes for enrichment analysis.The results showed that most regulated genes were significantly enriched in pathways related to the human immune process,such as the same species.Allogeneic rejection,graft versus host disease,intestinal immune network for the production of Ig A,autoimmune thyroid disease,Staphylococcus aureus infection,antigen processing and presentation,inflammatory bowel disease,etc. |
