| Purpose:Cardiovascular disease(CVD),especially coronary heart disease(CHD),remains one of the maj or causes of human death worldwide.However,the causes of CVD have yet to be fully understood through substantial progress in prevention and control.In recent years,much effort has been devoted to exploring and identifying new risk factors for atherosclerosis.In addition to genetic factors,environmental factors such as nutrition and gut microbiome are also considered to be important in the development of CVD.Nevertheless,the relationship between the gut microbiome,especially the virome,and CVD remains unclear.In this study,by analyzing and comparing the gut viral virome of patients with CHD and healthy adults,a background database of their viral viromes was constructed to provide data support for clarifying the relationship between gut viral virome and the occurrence and development of CHD in the future.Methods:1.Viral Metagenomic methods were used to analyze the composition of the gut viral virome of patients with CHD and healthy adults and to compare the differences between the two groups of viral virome by alpha and beta diversity analysis.2.Bioinformatics and molecular biology methods were used to analyze high-titer viruses in intestinal fecal samples of patients with CHD.Further,genetic characteristics of high-titer novel viruses in intestinal fecal samples of patients with CHD were explored by constructing a phylogenetic tree and analyzing genomic structure.3.SWISS-MODEL and MEME suite were used to predict the protein structure and function of the unique novel virus-conserved region protein in intestinal fecal samples of patients with CHD and to conduct an epidemiological investigation by polymerase chain reaction.Results:1.Analysis and comparison of gut virome:The gut viral virome of patients with CHD mainly comprised of phages and plant viruses,accounting for 54.68%and 43.25%,respectively.The gut viral virome of healthy adults was mainly composed of phages and vertebrate and invertebrate viruses,accounting for 71.31%and 28.15%,respectively.Comparing the two groups showed a significant difference in the diversity of viral species between gut viral viromes of patients with CHD and healthy adults.Healthy adults have a more decadent array of gut viral virome.The differences in the composition of the two viruses mainly include the following:Compared with healthy adults,the number of phages in the gut viral virome of patients with CHD was significantly decreased(35.26%/58.74%),and the number of Siphoviridae was mainly decreased(17.35%/35.86%).Inoviridae was found in the gut viral virome of patients with CHD but not in the gut viral virome of healthy adults.In addition to phages,plant viruses were dominant in the gut viral virome of patients with CHD.In contrast,vertebrate and invertebrate viruses dominated the gut viral virome of healthy adults.2.Identification and genetic characterization of phages in the gut viral virome of patients with CHD:44 Caudovirales virus sequences with complete terminal large subunit(TerL),21 complete virus sequences of Microviridae,and six complete virus sequences of Inoviridae were assembled from the CHD intestinal sample libraries.According to the virological classification criteria of ICTV and phylogenetic analysis,six strains of the Inoviridae virus were identified as a new virus genus.Phylogenetic analysis based on amino acid sequences of 44,21,and six virus-conserved region proteins showed high genetic diversity in intestinal fecal samples from patients with CHD.3.By predicting the conserved motifs and three-dimensional structure of Zot proteins of six strains of the novel Inoviridae virus,it was found that all Zot proteins of six strains contained conserved motifs expressing ATPase-like action and different amounts of unknown hypothesis protein motifs.Strains SHuInol-1,SHuInol-2,SHuIno3-2,SHuIno5-1,and SHuIno7-1 also contained conserved motifs encoding β-lactamase family proteins.The prediction of the three-dimensional structure of the Zot protein of six strains showed that the structural models of the Zot protein of SHuInol-2 and SHuIno5-1 were similar.In comparison,the three-dimensional structure models of the proteins of the other four strains were significantly different in topology.The epidemiological investigation showed that the favorable rates of six novel viruses in intestinal fecal samples of 43 patients with CHD were 2.33%,2.33%,4.65%,2.33%,2.33%,and 4.65%,respectively.The prevalence rates were all lower than 5%.Conclusion:This study analyzed the characteristics of the gut viral virome of patients with CHD.It established a background database of the gut viral virome of patients with CHD and healthy adults.The results showed that the diversity of the gut viral virome of patients with CHD was lower than that in healthy adults,the plant virus in the gut viral virome of patients with CHD was significantly more than that in healthy adults,the phage titer in the gut viral virome of patients with CHD was higher and had rich genetic diversity,and the prevalence of novel viruses was lower in the population with CHD.This study enriched the understanding of the characteristics of the gut viral virome of patients with CHD.It provided strong data support for the future,elucidating the relationship between the gut viral virome and the occurrence and development of CHD. |
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