The Characteristics Of Microbial Communities Of Oral Cavity,Esophagus And Cardia Of Population In High-risk Regions Of Esophageal Cancer In China

ObjectivesThis study aimed to describe and compare the characteristics and composition of microbiome of oral cavity,esophagus and cardia in Chinese population in high-risk regions of esophageal cancer,and to investigate the association between specific microbiota and esophageal squamous cell carcinoma(ESCC),gastric cardia adenocarcinoma(GCA),and dysplasia.The objective of this study was to add new microbial evidence to the etiology of esophageal and cardia cancer.Material and methodsThe characteristics of microbiome of patients with ESCC and GCA in high-risk regions of esophageal cancer in China.Tumor tissues and nontumor tissues from 67 patients with ESCC and 36 patients with GCA diagnosed in Esophageal Cancer Hospital in Linzhou,Henan province were collected.16S ribosomal RNA(16S rRNA)V4 gene region was sequenced using Illumina MiniSeq and processed using Quantitative Insights into Microbial Ecology(QIIME1).The microbial characteristics including a diversity(observed species and Shannon index),? diversity(weighted and unweighted Unifrac distance matrices),and relative abundance of taxa from taxonomic levels(phylum,class,order,family,and genus levels)were described.The linear discriminant analysis effect size(LEfSe)method was used to identify differentially abundant microbes,the Wilcoxon rank-sum test was used to test a diversity differences,and Permutational Multivariate Analysis of Variance(PERMANOVA)was used to test for differences in ? diversity.Oral microbiome of patients with esophageal and cardia diseases in high-risk regions of esophageal cancer in China.This study characterized the oral microbiome of 60 healthy people,97 patients with ESCC dysplasia,98 patients with GCA dysplasia from the screening cohort in Linzhou,Henan province and 110 patients diagnosed as ESCC and GCA at Esophageal Cancer Hospital in Linzhou,Henan province.Saliva,tongue swab,and dental plaque samples were collected,16S rRNA V4 gene region was sequenced using Illumina MiniSeq and processed using QIIME2.? diversity including observed species,Shannon index,and Faith's PD(F aith's phylogenetic diversity)were calculated.Microbial communities(Bray-Curtis,weighted and unweighted Unifrac distance matrices)of healthy people,patients with mild,moderate,and severe esophageal dysplasia,ESCC and patients with intestinal metaplasia(IM),low-grade intraepithelial neoplasia(LGIN),high-grade intraepithelial neoplasia(HGIN),GCA were compared using PERMANOVA.The association between specific taxa and ESCC or GCA was examined using multivariate logistic regression model and random forest model.Dynamic changes of oral microbiome of Chinese population in high-risk regions of esophageal cancer:a longitudinal study.Paired saliva samples were collected from 36 participants in screening cohort at two time points with the time interval of one year.16S rRNA V4 gene region was sequenced using Illumina MiniSeq and processed using QIIME2.Wilcoxon signed rank-sum test and Bonferroni correction were used to compare the changes of microbial communities.ResultsThe average ages for patients with ESCC and GCA were 62.2(SD,6.8 years)and 62.1(SD,6.6 years),respectively.In the GCA tumor tissues,the Shannon index(4.3 vs 2.0;P=0.001)and the observed species(102.5 vs 58.0;P=0.001)were significantly higher in comparison with the nontumor tissues..When we considered the microbial community composition(the ? diversity),significant clustering was detected for the weighted UniFrac distance between paired ESCC tumor and nontumor tissues(P=0.015).Paired GCA tumor and nontumor tissues showed 2 distinct clusters by tumor status in both weighted and unweighted UniFrac distances(P=0.004 and P=0.002,respectively).ESCC tumor tissues had greater relative abundances of Fusobacteria at the phylum level(3.9%vs 1.7%)with consistent associations at all taxonomic levels,including Fusobacterium at the genus level(3.2%vs 1.3%).GCA tumor tissues had higher relative abundances of Prevotella(23.3%vs 13.7%),Streptococcus(11.2%vs 4.2%),Veillonella(8.6%vs 3.7%),Haemophilus(4.3%vs 1.0%),and Neisseria(4.8%vs 1.5%)and lower relative abundance of the Helicobacter genus(11.8%vs 60.5%)in comparison with nontumor tissues.We found a positive association between the relative abundance of Fusobacterium and increasing tumor stage in ESCC tissues.Within patients with GCA,the relative abundance of Helicobacter(P=0.500)decreased and Prevotella increased(P=0.080)with a more advanced tumor stage.There were no statistically significant differences in ? diversity as measured by the Shannon index(4.3 vs 4.4)and observed species(83.0 vs 104.5)for ESCC and GCA tumor tissues;however,ESCC nontumor tissues had significantly increased a diversity as measured by the Shannon index(3.8 vs 2.0;P<0.001)and observed species(87.0 vs 58.0;P=0.001)in comparison with GCA nontumor tissues.Principal coordinate analysis plots based on the weighted and unweighted UniFrac distances suggested clustering of ESCC and GCA tumor tissues(P=0.017 and P=0.004,respectively).In addition,distinct clustering was detected for ESCC and GCA nontumor tissues as measured by both weighted and unweighted UniFrac distances(P=0.001 for both).Dental plaque had higher Shannon index than saliva and tongue(P<0.001),but saliva had the highest observed species(P<0.001).As for ? diversity,PCoA plots based on both Bray-Curtis and Unifrac distance matrices showed that the microbial communities of dental plaque were different from saliva and tongue.Different parts of oral cavity contributed 15.0%to the overall variance of oral microbiome.Patients with esophageal severe dysplasia had lower Faith's PD in saliva compared with healthy people after the adjustment of age,sex,smoking,drinking,education levels,BMI(Body Mass Index),fruit intake,and loose tooth(P=0.030).Patients with IM,LGIN,and GCA had higher observed species than healthy people,but patients with HGIN had marginally lower Faith's PD compared with healthy people after the adjustment of age,sex,smoking,drinking,and fruit intake(P=0.060).The relative abundance of Rothia was positively associated with ESCC dysplasia adjusted by age,sex,smoking,drinking,education levels,fruit intake,and loose tooth(OR=1.25,P=0.002 and OR=1.80,P<0.001 for mild and moderate dysplasia,respectively).The relative abundance of Capnocytophaga and Veillonella were associated with 1.67-and 1.30-times greater odds of ESCC(P<0.001).For GCA,the relative abundance of Rothia was positively associated with IM adjusted by age,sex,smoking,drinking,and fruit intake(OR=1.30,P<0.001).We compared the oral microbial communities between two samples before and after one year and found the diversity and relative abundance of taxa changed within individuals,but there was no statistical difference in general(P>0.050).Patients with regressed esophageal diseases had higher relative abundance of Prevotella than patients who were progressed within one year.The relative abundance of Neisseria,Porphyromonas and Rothia increased in patients with progressed esophageal diseases.ConclusionsThe microbial compositions of paired tumor and nontumor tissues were distinct for both patients with ESCC and patients with GCA.In ESCC tumor tissues,the relative abundance of Fusobacterium increased.In GCA tumor tissues,there was a large decrease in the relative abundance of Helicobacter.For nontumor tissues,the esophagus and the gastric cardia appeared to be distinct communities,but in the tumor,the microbial communities appeared to be slightly more similar.These findings are intriguing,but additional case-control studies as well as large-scale prospective cohorts are needed to verify our associations with specific taxa and ESCC or GCA.Oral microbial communities were different among saliva,tongue,and dental plaque.Patients with severe esophageal or cardia disease had different oral microbiome compared with patients with mild esophageal or cardia disease or healthy people.The relative abundance of Rothia in saliva was associated with increased risk of both esophageal and cardia dysplasia.Participants with more Capnocytophaga and Veillonella had higher risk for ESCC compared with participants with lower relative abundance of those two taxa.Some specific taxa may have potential role as biomarkers for esophageal and cardia cancer in the future.The microbial diversity and composition of saliva changed within individuals but with no significant differences in general.The changes of microbial communities in saliva over time should be taken into consideration for large-scale prospective studies in the future.