Multi-omics Analysis Of Pathogenesis And Immune Response Of Angiosarcoma And COVID-19

Background:The development of high-throughput technology has brought about tremendous changes in medical research.Multiple omics data across multiple cellular functions,provide unparalleled opportunities to understand the biology of complex diseases,such as cancer.Taking angiosarcoma(Angiosarcomaz)and new coronary pneumonia(Coronavirus disease2019,COVID-19)as examples,this paper aims to explore the application of multi omics technology in the study of complex diseases,and how to analyze the mechanism,diagnosis,prognosis and potential of gene level of diseases through omics data,so as to provide basis for the treatment of diseases.1 Whole genome and whole exome sequencing technology to study the molecular mechanism of angiosarcomaObjective:to analyze the genomic characteristics of angiosarcoma by whole genome sequencing and whole exome sequencing,and explore new biomarkers and new therapeutic targets of angiosarcoma.Methods:the clinical data of 47 patients with angiosarcoma were summarized through medical record and follow-up investigation;The total DNA of FFPE tumor samples was obtained and the whole genome and whole exome were sequenced;Quality control and bioinformatics analysis of the data.Results:a total of 2474 somatic mutations were identified by genomic analysis,with a mean of 112.5(1.875 per coding megabase)and a range of 14 to 912 mutations per sample.25 high-frequency mutant genes were found,suggesting the possibility that KDR mutation might sever as specific marker to discriminate angiosarcoma cases with and without malignant potential.The analysis of mutation characteristics showed that the defective DNA mismatch repair and aging may contribute to the development of angiosarcoma.Log-rank analyses revealed that 8 SCNA regions,including those involving the gain of 3q29,4q12,10p11.21,13q21.1,16p11.2.1,and17q12,and the loss of 7p22.3,and 9q34.3,were identified as significant predictors of poor progression-free survival.Conclusions:the combined analysis of whole genome and whole exome confirmed that there were widespread gene mutations in patients with as,revealed the correlation between somatic mutations and clinicopathological features,clarified the potential mechanism of the occurrence and development of angiosarcoma,and provided new insights to improve the treatment of angiosarcoma.2 Analysis of tumor immune microenvironment of angiosarcoma by multiple immunofluorescence techniqueObjective:to analyze the heterogeneous distribution of immune cells in the tumor microenvironment of angiosarcoma by multiple immunofluorescence technique,and to understand the role and function of each component in the tissue microenvironment,so as to predict the prognosis of immunotherapy more accurately.Methods:tumor tissue sections of47 patients with angiosarcoma were stained by multiple immunofluorescence staining.Pannoramic MIDI,Qupath and 3DHISTECH’s Slide Converter software were used for analysis.Tumor areas were delineated in sections and the average density of each immune cell in each patient was calculated by quantifying the immune cells and tumor area within the tumor area(num positive cells/mm^2).Graph Pad is for statistics and graphics.Results:Angiosarcoma is an immunosuppressive tumor microenvironment dominated by myeloid cells,and the cell density of tumor-associated macrophages(TAMs)is the highest in the microenvironment.In the part of lymphoid immune cells,the density of CD3~+T and CD8~+T cells is higher,and the abundance of T cells was associated with better survival in patients with angiosarcoma.Tumor infiltrating immune cells are mainly distributed near tumor blood vessels.In some patients,T cells and B cells in tumor will form tertiary lymphoid structure(TLS)around vascular endothelial cells,which is significantly correlated with the survival and prognosis of patients.Stem cell like CD8T cells(TCF1~+CD8~+T)in APC niche in tumor play an important role in tumor immunotherapy.Conclusions:multiple immunofluorescence technique can comprehensively analyze the tumor immune microenvironment of angiosarcoma at the protein expression level,so as to provide a reference basis for the selection of gene targeting or immunotherapy in the future.3 Transcriptomics and proteomics elucidate the mechanisms of lung injury and immune dysregulation in COVID-19Objective:integrating transcriptome and proteomics data sets to analyze the mechanism of lung injury and changes in the pulmonary immune system in COVID-19 patients,providing a basis for the pathogenesis of COVID-19 and the development of effective antiviral drugs.Methods:we obtained lung samples from nine patients who died of COVID-19 during the initial outbreak.HE staining was used to observe the pathological characteristics of lung tissue specimens from patients with COVID-19,and immunofluorescence was used to detect the expression of specific proteins in lung tissue.The sample RNA was extracted and sequenced to analyze the gene expression changes caused by SARS-COV-2 infection in the lung tissue of COVID-19 patients.Protein was extracted and digested from lung tissue samples,and detected by LC-MS/MS technology.Proteomic information of lung samples from COVID-19 patients was analyzed.Results:all patients had histological evidence of diffuse alveolar damage,with widespread hyaline membrane formation,evidence of fibrosis,and varying degrees of an inflammatory infiltrate.Notably,several patients were noted to have a significant number of neutrophils present in the lung tissue.we show that pathways related to neutrophil activation and pulmonary fibrosis are among the major up-regulated transcriptional signatures in lung tissue obtained from COVID-19 patients.Strikingly,the viral burden was low in all samples,which suggests that the patient deaths may be related to the host response(such as inflammatory response)rather than an active fulminant infection.Proteomics analysis validated our transcriptome findings and identified several key proteins,such as the SARS-Co V-2 entry-associated protease cathepsins B and L and the inflammatory response modulator S100A8/A9,that are highly expressed in fatal cases.Conclusions:lung transcriptome and proteomic analysis of 9COVID-19 patients who in the early stage of the epidemic elucidated the mechanism of lung injury caused by COVID-19,elucidated the causes of pulmonary immune dysfunction in COVID-19 patients,and provided potential therapeutic targets for patients.