RNA-Seq Explores The Transcription Landscape Of Atherosclerotic Platelets And The Interaction Of Key Genes With Dendritic Cells

Objective: Platelet genome sequencing was carried out in patients with acute coronary syndrome and healthy physical examination by transcriptome sequencing(RNA-Seq)technology.m RNA of platelet differential expression genes was screened out,key genes were searched for analysis,and the receptor of this molecule on dendritic cells was explored to verify whether there was interaction between platelets and dendritic cells in the process of atherosclerosis.It provides a new research direction for early diagnosis and treatment of patients.Method: 1.Peripheral blood of 6 patients with acute coronary syndrome and 6 healthy physical examination patients admitted to the Department of Cardiology,Affiliated Hospital of Qingdao University from December 2020 to January 2021 were collected,and m RNA sequencing was performed by transcriptional sequencing(RNA-Seq)technology to analyze the genetic differences.GO enrichment analysis and KEGG signaling pathway analysis related to differentially expressed genes were sought.2.In addition,peripheral blood platelets of 20 patients with ACS and 20 patients with healthy physical examination were collected,and the highest differentially expressed genes were verified by q RT-PCR.The gene CCL3 with the highest expression was selected from the differentially expressed genes,and the differential expression of platelet CCL3 in ACS patients and healthy subjects was detected by Western Blot,and the expression level of CCL3 in peripheral blood of the two groups was detected by Elisa.3.The expression of CCR5,the receptor of CCL3,on dendritic cells was verified by co-immunoprecipitation method.Results: A total of 815 differentially expressed mrnas were screened from patients with ACS compared with healthy subjects,among which 524 genes were up-regulated and 291 genes were down-regulated.GO enrichment analysis and KEGG pathway analysis were performed to find the enrichment and pathway related to platelet chemotaxis,adhesion and aggregation.GO enrichment analysis found that differentially expressed m RNA participated in biological processes such as metabolism,extracellular matrix,immune response and inflammatory response.The differentially expressed genes of platelet up-regulation are mainly concentrated in neutrophil-mediated immunity,granulocyte activation,neutrophil-degranulocyte activation,antimicrobial humoral response,angiotensin maturation,regulation of blood angiotensin level,regulation of angiotensin metabolism,humoral immune response,regulation of peptidase activity,and regulation of endopeptidase activity.In KEGG signaling pathway analysis,FC-γ receptor signaling pathway was involved in phagocytosis,protein processing regulation,BP protein maturation regulation,CC antigen binding,homologous recombination,lipid metabolism,renin secretion,and FC-γr-mediated phagocytosis.Further,q PCR technology was used to verify that the genes with the highest differential expression were up-regulated top10 genes: CCL3 OLAH GPIHBP1 H19 RARRES2P11 AC002044.2AREG ADGRL4 and down-regulated Top 10 genes: GAPDHP61 INTS6P1 FRAS1 SLC7A3 IGKV1-8PRINK1 MRPL3P1 GPC3 SHISA2 NXF3,Gene related to platelet activation: IGF2COL1A2 ORM1 TIMP3 GP9 TIMP1 ITGA2 B SERPING1 F8 ALDOA PLA2G4 B A1BG,and the gene CCL3 with the highest differential expression was found.Western Blot and Elisa were used to verify that the differential expression of CCL3 was also statistically significant.In the comparison of general information,only c Tn I and NT-Pro BNP had statistical significance,and there was no statistical difference in the age and gender of patients.The expression of CCR5,a receptor containing CCL3,on dendritic cells was verified.Conclusion: RNA-Seq was used to screen out a large number of genes with differential expression in platelets between ACS patients and healthy subjects,among which the gene with the highest expression was related to chemotaxis,adhesion and aggregation,while CCL3 with the highest differential expression might mediate the interaction between platelets and dendritic cells by affecting the receptor CCR5 molecule of dendritic cells.Involved in the process of atherosclerosis.