Gene Expression Changes Of Peripheral Blood Mononuclear Cells In Primary Immune Thrombocytopenia Were Investigated By MRNA Sequencing

Objective: To use mRNA sequencing and high-throughput sequencing technology to sequence peripheral blood mononuclear cells(PBMC)from patients with primary immune thrombocytopenia(ITP)and healthy individuals,obtain mRNA expression information from each group,and systematically analyze changes in mRNA expression in the PBMC of newly diagnosed ITP patients.The study aims to verify mRNA that is significantly associated with the disease using qPCR,explore mRNA closely related to the occurrence and development of ITP,provide theoretical basis for the role of mRNA in the pathogenesis of ITP,explore potential immune mechanisms related to the occurrence and development of ITP,and discover potential target genes.Method: High-throughput mRNA sequencing was performed on PBMCs from ITP patients(n=4)and healthy adults(n=4).The identification of differentially expressed genes(DEGs)between the groups was performed using R language computer information technology with Pvalue<0.05 and|log2Foldchange| ≥ 1 as the criteria for DEGs.The DEGs were annotated using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The protein interaction network of the DEGs was constructed using the STRING online database,and hubgenes were identified using Cytoscape software and the cyto Hubba plugin(Dgree algorithm).Key genes were validated using real-time fluorescent quantitative PCR(qPCR).Results: Comparison of mRNA expression data between ITP patients and healthy controls led to the identification of 1,291 DEGs,of which 785 were up-regulated and 506 were down-regulated.GO enrichment analysis showed that the biological processes(BP)were enriched in inflammatory response,immune response,innate immune response,and phagocytosis;the DEGs of the cellular components(CC)were mainly enriched in phagolysosomes,extracellular exosomes,plasma membranes,and specific granules;and the molecular functions(MF)were mainly enriched in RAGE receptor binding,protein binding,Ig G binding,and arachidonic acid binding.KEGG pathway enrichment analysis showed that DEGs were mainly enriched in Toll-like receptor signaling pathway,Fc gamma R-mediated phagocytosis,and natural killer cell-mediated cytotoxicity.The protein interaction network and Cytoscape software visualization analysis revealed that MAPK14,MAPK3,TNF,MYD88,and IL-1β were highly connected hubgenes.The qPCR results showed that compared with the healthy control group,the mRNA expression of MAPK14,MAPK3,TNF,and MYD88 genes in the ITP group was significantly increased(P<0.05),and the expression of IL-1β was slightly higher than that of normal individuals,but it was not statistically significant(P=0.051).Conclusion: Analysis of mRNA sequencing data from PBMCs of ITP patients and healthy controls suggests that the MYD88-dependent Toll-like receptor signaling pathway may be a key signaling pathway for the development of ITP,and the transcription levels of related target genes,including MAPK14,MAPK3,TNF,MYD88,and IL-1β,are abnormally altered in the PBMCs of ITP patients.The results of this study provide a theoretical basis and new directions for investigating the immunological pathogenesis and exploring diagnostic and therapeutic targets for ITP.