人体皮肤菌群的培养组学方法建立和治疗特应性皮炎的菌种资源挖掘

近年来，国内外学者通过宏基因组技术获得了大量人体疾病和微生物关联的信息，但纯培养仍然是感染性疾病研究的基础。目前，肠道菌群培养组学和菌群移植治疗疾病，已取得突破性进展，但在皮肤菌群方面却鲜有报道。特应性皮炎(Atopic dermatitis, AD)是一种典型的与菌群多样性密切相关的皮肤疾病，其患者皮肤存在严重的菌群失调，且恢复菌群多样性有利于疾病的改善和恢复，从而为微生物相互作用和菌群移植治疗AD提供理论依据。本项目拟以人体皮肤菌群为研究对象，在过去多年研究特殊生境微生物分离方法的基础之上，结合质谱和16SrRNA、ITS基因测序，从而建立皮肤菌群的培养组学方法；利用宏基因组分析，探索潜在关键菌种；通过对关键菌种的培养组、代谢组、网络等分析，深入解析其“株”水平上的功能差异及其代谢产物的变化规律，结合实验验证，筛选治疗AD的关键菌株并探讨其功能，从而为皮肤疾病的菌群移植疗法奠定基础。

The development of omics technologies by researchers at home and abroad, beginning with metagenomics, highlighted the role of gut flora in several disorders, while bacterial cultures still play important role in the diagnosis and treatment of pathogenic bacteria of current infectious diseases. At present, breakthrough progress has been made in the culturomics for human gut microbiota, and intestinal microbiota transplantation (Fecal Bacteriotherapy) for recurrent Clostridium difficile infection. There are, however, limited reports about the isolation and culture techniques of human skin microbiota, and microbiota transplantation for treating skin disease in China. Atopic dermatitis (AD) is a chronic, relapsing, and pruritic skin disease, characterized with typical skin microbiome dysbiosis. Both 16S rRNA and metagenomic analyses of bacterial DNA have shown that the skin microbiome on AD subjects has decreased bacterial diversity and increased Staphylococcus aureus colonization. The dysbiosis in AD is critical to driving disease severity and/or tendency for relapse. Additional, the flora diversity restoring is beneficial to the AD’s recovery. These observations support the hypothesis that skin microbiota transplantation can be used to treating AD. This project plans to do an extensive study on human skin flora and aims to establish a series of selective isolation methods on the basis of matrix-assisted laser desorption/ ionization time of flight and 16S rRNA gene, ITS gene for identification experience of isolation of microbes living in those unusual environments over the past years. Potential key strains would be accessed by metagenomics studies; functional genes differences and their metabolites at the "strain" level will be analyzed by culturomics, genomics, and network analyses. The key stains will be further verified by in vitro experiments. This project intends to solve the problem about the culturomics of human skin flora, and explore those key strains and functional genes related to skin diseases, thereby revealing the mode of interactions between host and microorganisms, and between different microorganisms. Meanwhile, it would help researchers to confirm that interventions targeting the microbiome and novel antibiotic impair pathogen colonization could provide therapeutic benefit for patients with AD. Furthermore, the culturomics may provide new insights into diagnostics and personalized medicine.