The Lung Microbiome In Respiratory Diseases-Next Generation Sequencing Technology

Background:In clinical practice,treatment was stepped up to antibiotic treatment and "triple therapy"(ICS,LAB A and LAMA)in acute exacerbations of chronic obstructive pulmonary disease(COPD),but some of COPD patients seem to be corticosteroid-insensitive or antibiotic-resistant.Recently,the use of antibiotics is thought to influence the balance of bacterial and fungal community(mycobiota),leading to the resistance to antibiotic.However,few studies had characterized the fungal community dynamics in AECOPD patients.Moreover,COPD is the risk factor of lung cancer,some of the reaches has studied about the if COPD and lung cancer share the same risk factors.Recently,increasing evidences have proved lung microbiome play an important role in COPD,some of study has focus on lung microbiome and lung cancer.However,study focused on lung microbiome in patient with lung cancer and COPD comorbidity.Objective:To test the hypothesis that the composition of lung mycobiome might affect the therapeutic efficacy of antibiotics and "triple therapy" and the use of antibiotic might change the structure of the lung mycobiome instantly.Methods:We collected a total of 44 sputum samples from 44 AECOPD patients at the first hospitalized day and identified bacterial and fungal species by using 16S rRNA and ITS sequencing.We characterized and compared the lung mycobiome in AECOPD patients with and without improving dyspnea after antibiotic and "triple therapy",with and without antibiotic treatment.We also collected a total of 49 bronchoalveolar lavage fluid(BALF)of patients with lung cancer and 7 BALF of patients with benign mass,as well as 4 control cases before fiberoptic bronchoscopy.We characterized and compared the lung microbiome between patients with lung cancer and patients with benign mass,between patients with adenocarcinoma and squamous cell carcinoma.Results:We found that the most common fungal population in AECOPD patients were the phylum ascomycota(57.20%)including sordariomycetes Other,aspergillus,phialemoniopsis,candida,trichocomaceae_Other,and phylum basidiomycota(14.97%)including schizophyllum,heterochaete,trechispora,cystofilobasidiales Other,agaricomycetes Other and filobasidiales_Other.In the subgroup of patients without antibiotic treatment in the prior 1 week,significant decreases in fungal alpha diversity(PD_Whole Tree,p=0.008)and increases in the abundance of phylum ascomycota(p=0.000)were observed in patients without improving dyspnea,compared with the controls,while the bacterial communities were much more similar.In the subgroup of patients with improving dyspnea,fungal communities in patients with antibiotic treatment in the prior 1 week differed significantly from the controls.Phylum ascomycota was significantly more abundant(p=0.000)in patient with antibiotic therapy than the controls,while basidiomycota was more abundant(p=0.002)in the controls,and the significant decreases in fungal alpha diversity(PD_Whole_Tree,p=0.000)were observed in patients with antibiotic therapy compared with the controls.We found that the most common bacterial population in lung cancer patients were the phylum Proteobacteria(47.63%),Firmicutes(25.14%),Bacteroidetes(17.91%),Actinobacteria(4.34%),Fusobacteria(2.21%)and Cyanobacteria(1.32%).In the genus level,the most common bacterial population included Halomonas,Streptococcus,Prevotella,Veillonella,Ralstonia,Neisseria,Haemophilus,Fusobacterium,Pseudomonas,Porphyromonas,Acinetobacter,Aggregatibacter,Achromobacter,Lactococcus and Nesterenkonia.Although the alpha diversity between the groups has no significant difference,we had found that adenocarcinoma and squamous cell carinoma have own special bacteria.In the Group Aden,bacteria that were significantly different from other groups were Pasteurellaceae and Pasteurellales.In the Group SQC,bacteria different fromm other groups were bacteroidetes,Streptococcus,Bacteroidales,Bacteroidia,Prebotellaceae,Prevotella,Fusobacteriales,Fusobacteria,Fusobacterium,Fusobacteriaceae,Porphyromonadaceae,Porphyromonas,Flavobacteriaceae and Capnocytophaga.Conclusions:Our study showed significant alternations in lung mycobiome with improving dyspnea and with antibiotic treatment for COPD.The composition of lung mycobiome affects the therapeutic effcicay of antibiotic and "triple therapy" as well as that the antibiotic treatment changes the structure of the lung mycobiome instantly.Moreover,ascomycota might be one of the prognostic markers in AECOPD.Although the alpha diversity between the group Aden and group SQC has no significant difference,adenocarcinoma and squamous cell carinoma have their own special bacteria.Moerover,the lung microbiome compositions were different between the patients with AECOPD and with lung cancer.