Exploratory Analysis Of Chromatin Accessibility And Related Transcription Factors In Lung Immune Cells Of Sepsis Mouse By Single Cell Assay For Transposase Accessible Chromatin Sequencing

Objective:Recently,the mechanism of sepsis-induced immune cell dysfunction in lung tissue is not clear,and this study used single cell assay for transposase accessible chromatin sequencing as the entry point to analyze the chromatin accessibility of immune cells at the genome-wide level,explore relevant pathogenic genes,pathways and biological processes,and find related transcription factors to provide new ideas for the targeted therapy of sepsis-induced lung injury.Method:A mouse model of sepsis was first established by cecal ligation and puncture and a single-cell suspension was prepared,Mouse lung tissue CD45~+ cells were obtained by flow cell sorting,followed by single cell ATAC sequencing of this types.Cell subsets were clustered using Loupe Cell Browser and obtained differential chromatin opening for each cell subset.Pathway and biological process analysis were performed through the Metascape online analysis tool.In addition transcription factor accessibility region analysis of Loupe Cell Browser was used to search for differential transcription factors in each cell subpopulation.Finally,experimental validation of differential transcription factor ZBTB7 B,the expression of transcription factor ZBTB7 B in CD3~+ T lymphocytes from lung tissue of septic mice and normal healthy mice,and in CD3~+ T lymphocytes from the peripheral blood of septic patients and normal healthy humans.Results: CD4~+ T lymphocytes,neutrophils,and NK cells in CD45~+ immune cells of CLP-induced septic mice were significantly increased relative to controls.Their inflammation-related pathways and biological processes were activated,the expression of genes related to pathways and biological processes was increased,and the inflammatory biological processes were interrelated between subsets of cells and subsets.The transcription factor ZBTB7 B was increased in CD4~+ T lymphocytes and in CD8~+ T lymphocytes,and the transcription factor ZBTB7 B was increased in total T lymphocytes(CD3~+ T lymphocytes)and CD3~+ T lymphocytes in septic patients.Conclusion:After sepsis causes lung injury,the level of chromatin opening in lung tissue is increased,inflammation-related pathways and biological processes are activated,and the expression of genes involved in pathways and biological processes is increased.And that the transcription factor ZBTB7 B is involved in the differentiation process of T lymphocytes to CD4~+ T lymphocytes during septic lung injury.