Exploring The Molecular Biological Network Of PrimarySj(?)gren’s Syndrome And Potential Small Molecule Drugs Based On A Comprehensive Analysis Of Transcriptomic And Gene-wide Association Studies

Objective:To explore the pathogenesis and potential small molecule drugs of primary Sjogren’s syndrome(pSS)based on comprehensive analysis of transcriptomics and gene-wide association studies.Methods:The following sentence was translated using the GEO database to retrieve datasets of bulk RNA-seq and single-cell RNA-seq from peripheral blood mononuclear cells(PBMC)and salivary glands from patients with primary Sjogren’s syndrome(pSS).The datasets were quality controlled using the limma package in R software,and differential expression genes were selected using the clusterProfiler,MCODE,and WGCNA packages for systems biology enrichment analysis and visualization of relevant molecular mechanisms.The xCell,CIBERSORTx,and ImmuneCellAI packages were used for immune cell infiltration analysis of PBMC and salivary gland bulk RNA-seq data from pSS patients.Single-cell RNA-seq data from pSS PBMC were analyzed using Cellranger and Seurat4.0 to identify and draw a map of immune cells in pSS,and differential gene expression and cell-cell interaction analysis were performed on each immune cell subpopulation.The pSS whole-genome association study data and interferon(Ⅰ/Ⅱ)gene sets were further integrated for systems biology analysis and visualization,and small molecule drug prediction was performed using the cMAP package in R software.Results:Enrichment analysis of differentially expressed genes in PBMC Bulk-seq revealed that the pathogenesis of pSS may involve molecular mechanisms such as interferon-mediated signal transduction regulation,cytokine signal transduction in the immune system,IFN-mediated antiviral response,innate immune response pathway,MDA-5 signaling pathway,regulation of type Ⅰ interferon production,positive regulation of immune response,regulation of response to cytokine stimulation,regulation of tumor necrosis factor production,and regulation of immune cell response to cytokine stimulation.MCODE and WGCNA analysis showed that the biological molecular modules of PBMC immune disorder in pSS involve multiple processes of innate immune pathways and adaptive immune pathways,such as processing and presentation of exogenous and endogenous antigens,regulation of lymphocyte activation,adaptive immune response,regulation of T cell activation,immune mediated by lymphocytes and B cells,Th17 cell differentiation,and cytokine signal in the immune system.The immune cell infiltration analysis of PBMC Bulk-seq showed that the proportions of DC,B cell.Macrophage.Effector memory T,and Thl immune cells were significantly upregulated in the peripheral blood immune disorder of pSS.while the proportions of NK,NKT,MAIT,Th17,nTreg,and γδT immune cells were significantly downregulated.Enrichment analysis of differentially expressed genes in salivary gland Bulk-seq indicated that the mechanism of glandular damage in pSS may involve molecular mechanisms such as regulation of immune effector processes,leukocyte-mediated cytotoxicity regulation,humoral immune response,regulation of lymphocyte activation,positive regulation of immune effector processes,antigen processing and presentation,regulation of cell killing,adaptive immune response,cytokine-cytokine receptor interaction,and cell adhesion molecules.MCODE and WGCNA analysis showed that the biological molecular modules of salivary gland immune damage in pSS include biological processes such as processing and presentation of exogenous peptide antigens,processing and presentation of polysaccharide antigens,regulation of lymphocyte activation,adaptive immune response,antigen processing and expression of endogenous antigens,immune effector processes mediated by lymphocytes(T/B cells),immunoglobulin-mediated immune response,regulation of immune effector processes,interferon signaling pathway,cytokine signal in the immune system,ER-phagosome pathway,and antigen processing-cross presentation.The immune cell infiltration analysis of salivary gland Bulk-seq showed that the proportions of B cells.Memory B cells,CD4 memory T cells,CD8 Tcm,CD8 naive T cells,DC,pDC,Plasma,Tfh,γδT,and cDC immune cells were significantly increased.ThescRNA-seq data of PBMCs from pSS patients were divided into 20 clusters:Activated B cells,Activated NK cells,C11_DC(mesenchymal cell-like),C15_DC(SDPR+megakaryocyte;),C19_DC(PRSS57+stem cell),CD14 Monocyte,CD 16 Monocyte,CD3-CD4-CD8+T,CD4 Treg,CD8 T Effector,CD8 T Naive,Erythrocytes,Innate like T cells,Megakaryocytes,NELL2 CD8 T cells,pDC,Plasma B cells,Resting NK cells,Th17 cells,and γδ T cells.Cell proportion analysis revealed an increase in Th17 cells,CD8 T Effector cells,CD3-CD4-CD8+T cells,Activated B cells,Activated NK cells,Resting NK cells,CD14 Monocytes,and CD16 Monocytes in pSS.while CD8 T Na(?)ve cells,γδT cells.Plasma B cells,pDC,and Megakaryocytes were decreased.Further differential gene analysis of the 20 immune cell clusters in pSS pathology revealed molecular transcriptional regulatory mechanisms.The cMAP package was used to screen for candidate small molecule drugs based on the differential gene expression of the 20 immune cell clusters.Additionally.GWAS and single-cell analysis were integrated to capture subgroups of cells with potential genetic susceptibility(such as Th17 cells,Activated B cells,Monocytes,etc.),and some genetic susceptibility genes(such as TNF,OAS1,IFIT3,IFI27,IFI44L,TNFAIP3,CCR5,IRF5,etc.)were widely expressed in different cell types.Furthermore,combining type Ⅰ/Ⅱ interferon gene sets revealed that immune cell subgroups closely related to type Ⅰ interferon gene sets include Activated B cells,Monocytes,DC,pDC,Th17 cells,etc.revealing the pathogenic molecular mechanism of pSS.The interaction network between immune cells revealed that the communication between the 20 immune cell clusters in pSS PBMCs promoted immune damage through various ligand receptors or signaling pathways,further revealing the potential mechanism of mutual activation and proliferation of immune cells in pSS By merging and clustering T,B,NK,and Monocyte cell clusters separately and re-analyzing,more detailed immune cell subgroups were identified,providing new directions for understanding the pathogenesis of pSS.Conclusion:Based on the comprehensive analysis of the transcriptomic data of peripheral blood mononuclear cells(PBMC)and salivary glands in primary Sjogren’s syndrome(pSS),as well as the genome-wide association study data,the pathogenesis of pSS mainly involves type Ⅰ/Ⅱ interferons,immune cell cytokines and their mediated signal transduction pathways,innate immune pathways,and adaptive immune pathways related to immune cell activation,proliferation,and differentiation.Further analysis of immune cell infiltration emphasizes the increased proportion of innate immune cells(such as DCs,macrophages)and adaptive immune cells(plasma cells,Tfh,B cells,effector memory T cells)in peripheral blood and salivary glands of pSS.PBMC single-cell RNA-seq further characterizes the immune dysregulation mechanisms in pSS at a single-cell resolution.Combining with pSS GWAS data and type Ⅰ/Ⅱ interferon gene sets,the potential immune cell subsets and interferon-related pathogenic subsets of pSS’s genetic susceptibility are revealed.Cellchat analysis describes the communication among 20 immune cell groups in pSS PBMCs through various ligand-receptor pairs or signaling pathways to promote immune damage in pSS.The potential small-molecule drug comprehensive strategy explores potential small-molecule drugs for each cell subset,providing new strategies and directions for future drug screening and development for pSS.Objective:To explore the cellular map and molecular network mechanism of iguratimod(IGU)intervention in primary sjogren syndrome(PSS)based on single cell transcriptomics and system pharmacology.Methods:Firstly,the potential targets of IGU and PSS genes were predicted through the database,and the IGU-PSS targets were obtained by taking the intersection.Then,IGU-PSS targets were imported into Metascape for enrichment analysis,and related networks were constructed.Subsequently,the core targets of IGU-PSS targets were mapped to PBMC of PSS to find potential cell populations for IGU therapy.Finally,the molecular docking is used to verify the degree of binding stability of the IGU.Results:A total of 292 IGU and 524 PSS were obtained.The results of the IGU-PSS PPI found that the targets of IGU’s mainly intervention are AKT1,EGFR,MMP9,CASP3,CCL5,IL2,Serpinal,Elane,STAT1,CTSG,SELP,LCN2,NOS2,S100A9,MIF,CYP19A1,CYP19A1,VDR,LYZ.Further molecular docking results show that IGU and PSS’s AKT1,EGFR,MMP9,CASP3,CCL5,IL2 and other target docking kisses are better(less than-5 kCal/mol).Conclusion:IGU treatment PSS has the characteristics of multi-system,multi-component,and multi-target.Its possible mechanisms include regulating immunity,improving the proportion of lymphocyte sub-clusters,reducing the substrate of the gland,and blister damage,the proliferation,differentiation,and apoptosis of cells,and finally achieved the goal of controlling the progress of the disease.Objective:To evaluate the efficacy and safety of mesenchymal stem cell(MSC)transplantation in the treatment of autoimmune diseases.Methods:The Chinese and English databases were searched for clinical research on the treatment of autoimmune diseases with mesenchymal stem cells.The search time range is from a self-built database to October 1.2021.Two reviewers independently screened the literature according to the inclusion and exclusion criteria,extracted data,and evaluated the bias of the included studies.RevMan 5.3 analysis software was used for meta-analysis.Results:A total of 18 RCTs involving 5 autoimmune diseases were included.The 5 autoimmune disease were rheumatoid arthritis(RA),systemic lupus erythematosus(SLE),inflammatory bowel disease,ankylosing spondylitis,and multiple sclerosis.For RA,the current randomized controlled trials(RCTs)still believe that stem cell transplantation may reduce disease activity,improve the clinical symptoms(such as DAS28),and the percentage of CD4+CD 25+Foxp3+Tregs in the response group increased and the percentage of CD4+IL-17A+Th17 cells decreased.The total clinical effective rate of RA is 54%.For SLE,the results showed that mesenchymal stem cell transplantation may improve SLEDAI[-2.18(-3.62,-0.75),P=0.003],urine protein[-0.93(-1.04,-0.81),P<0.00001],and complement C3[0.31(0.19,0.42),P<0.00001].For inflammatory bowel disease,the results showed that mesenchymal stem cell transplantation may improve clinical efficacy[2.50(1.07,5.84),P=0.03].For ankylosing spondylitis,MSC treatment for 6 months may increase the total effective rate;reduce erythrocyte sedimentation rate,intercellular adhesion molecules,and serum TNF-α;and improve pain and activity.For multiple sclerosis,the current research results are still controversial,so more RCTs are needed to amend or confirm the conclusions.No obvious adverse events of mesenchymal stem cell transplantation were found in all RCTs.Conclusion:MSCs have a certain effect on different autoimmune diseases,but more RCTs are needed to further modify or confirm the conclusion.