Multi Omics Approaches To Study Interactions Between Human Adenovirus Type 55 And Host Cells

Human adenovirus is one of the common respiratory pathogens.In recent years,the respiratory tract infection caused by adenovirus is growing in China,and it is easy to cause outbreaks.Many outbreaks of adenovirus infection have been happened in army and schools,which has seriously affected the health of the Chinese.Studies have shown that the mortality of pneumonia caused by adenovirus infection can be as high as 50%in immunodeficient patients without treatment timely.Moreover,the detection rate of adenovirus in non-immune-deficient patients is also on the rise.In China,adenovirus has not been included in the list of respiratory pathogens,and there is no specific medicine for adenovirus infection.The existing adenovirus vaccines in the United States mainly target at type 4 and type 7,and they are not broad-spectrum.As a recombinant adenovirus,type 55 adenovirus has strong proliferation,and its infection causes a higher proportion of patients with severe disease.Nowadays,the spread of type 55 adenovirus regional characteristics,and it has been outbreaks in many places of China.At present,the severe mechanism of type 55 adenovirus infection is still unclear.Therefore,in-depth study on the interaction between adenovirus and host can provide potential drug targets and improve the treatment ability of adenovirus infection.In this study,a 55-type adenovirus isolated from an outbreak in a training center was used as the research object.A virus-host interaction model was constructed based on the adenovirus-susceptible human A549 cell line.Multiple infection time points were analyzed by dynamics transcriptome.Hi-C(High-through chromosome conformation capture)technology was used for three-dimensional conformation analysis to capture three-dimensional data of the interaction between HAd V55 and host cells.Comprehensive multi-omics data,RNA interference was used to identify the potential interaction gene sites between adenovirus type 55 and host cells.The results found that adenovirus type 55 began to transcribe and express after 24 h infection in A549 cells.And the transcription and expression of adenovirus from 72 h to96h increased exponentially.Through cluster analysis of differential expression patterns,it is found that the host cell differential genes caused by virus infection mainly gather in the three time periods of 6-24 h,48-72 h and 96 h,which are consistent with the time points of adenovirus transcription and expression in host cells.Significantly different genes in host cells at 6 time points from 6h to 96 h were 464,740,401,673,511,and808,respectively.Combined analysis of the 10 functional genes annotated by GO function and the 10 signal pathways enriched by KEGG found that significantly different host genes with adenovirus infection are mainly related to the positive regulation of cell growth,the promotion of cell apoptosis and antiviral infection-related genes.Through further analysis of the specific functions of the genes,eight related genes were identified,they are ARHGEF7,MAPK10,STK4,ABL2,GREB1,TANK,TRIM2 and TRIM24,respectively.According to the three-dimensional conformation capture technology,55-type adenovirus infection has changed the distribution of A/B compartments.Among them,the A/B compartment of chromosome 15 changed significantly,especially in the late stage of infection(72h and 96h).The distribution of adenovirus genome in A/B compartments showed that adenovirus genome was preferentially distributed in A compartment with higher transcriptional activity during the whole process of infection.Based on the transcriptome data,two host genes TJP1 and FANCI located on chromosome 15 were up-regulated after adenovirus infection.Combined with multi-omics data,the interaction sequences of these 10 genes(ARHGEF7,MAPK10,STK4,ABL2,GREB1,TANK,TRIM2,TRIM24,TJP1 and FANCI)with adenovirus genome were captured.It was found that gene ARHGEF7,GREB1,ABL2,TJP1,TRIM2 and TRIM24 interacted with adenovirus preferentially at 24 h.In the three time points from 48 h to 96 h,the interactions of 10 genes with the adenovirus genome were detected in varying degrees.Based on the multi-omics data,the relative changes of target genes and adenovirus after adenovirus-infected were detected after the down-regulated expression of 10 target genes by RNA interference.The results showed that when the expression of the gene FANCI with function of DNA repair,the gene STK4 with pro-apoptosis function,the gene TRIM24 with ubiquitination function,and the gene MAPK10 involved regulation of host inflammation were down-regulated,the load of adenovirus significantly increased.In addition,adenovirus infection increased the expression of gene ABL2,which promoted cell proliferation,also significantly increased the adenovirus load in cells.The results of multi-omics data showed the ten adenovirus genes,E1 A,E1B,L433 K 14.6 k Da protein,L4 p VIII 25 k Da protein,L2 protein V precursor,L2 p X 5.2 k Da protein,p IX,L4 100 k Da hexon protein,E3 12.2 k Da protein and L1,preferentially change during the process of interaction.Moreover,the interactions between the adenovirus genome and mitochondrial DNA of the host cell from 24 h to 96 h were first observed.The mitochondrial genes in the process of interaction were mainly energy transfer related genes ND1,ND5,ATP6,CO1 and CYB.In summary,this study used multi-omics techniques to display an in-depth analysis of the interaction between pathogenic adenovirus type 55 isolated from epidemic samples and host cells.The data has revealed the potential genes involved in the interaction between virus and host cell.Combined with RNA interference technology,the results showed that ABL2,TRIM24,FANCI,STK4 and MAPK10 were related to adenovirus infection.The gene ABL2 helps the replication of adenovirus in host cells through self-expression,TRIM24 and FANCI participate in the ubiquitination mechanism and DNA repair mechanism to resist adenovirus infections,while the genes STK4 and MAPK10 inhibit adenovirus transcription and infection by stimulating the pro-apoptotic mechanism and inducing inflammation.These gene targets establish a foundation for further understanding of the severe mechanism of adenovirus type 55,and provide an important reference for the clinical treatment of adenovirus infection and vaccine development.