The microbiome associated with oral Leukoplakia: A multi-omics mechanistic study

项目摘要 人们越来越关注微生物组在各种疾病的发展和/或进展中的作用。 癌症，包括口腔鳞状细胞癌（OSCC）。研究表明，口腔鳞状细胞癌与 独特的微生物组然而，OSCC本身代表了疾病过程的晚期阶段，其中观察到的 微生物的变化很可能是乘客或最好的情况下，晚驾驶事件。为了更好地理解 口腔鳞状细胞癌中的微生物组，并确定微生物生物标志物进行早期检测和预防，这是至关重要的， 评估疾病早期阶段的微生物组，例如口腔潜在恶性疾病（OPMD）。口服 白斑病（OL;本研究的重点）是最常见的OPMD。对微生物组知之甚少 与OPMD相关，其在其进展中的作用，或其作为非侵入性诊断生物标志物的潜力， 补充口腔上皮异型增生分级，目前最好的可用指标。我们的长期目标是 确定OPMD的诊断/预后微生物生物标志物，并确定微生物组对 口腔癌的发病机制进行了大规模的，国际财团研究分两个阶段：一个病例对照 研究（本申请），以全面研究与OL/发育不良相关的微生物群， 北美人群;随后是一项纵向研究（未来应用），将涉及随访 用于鉴定/验证恶性转化的微生物组生物标志物的相同和额外OPMD队列 （MT）。我们的假设是，与OL相关的微生物组组成和功能的变化代表了早期 与异型增生的严重程度相关的驱动事件，并有助于口腔癌的进展。在 此外，我们还假设口腔微生物的致癌菌株可能导致这些疾病。 为了解决这些假设，我们提出了3个具体目标：1）表征多王国的组成 与OL相关的微生物组和不同级别的发育不良; 2）定义微生物和宿主 OL中的转录组以及元转录组的这两个片段之间的潜在相互作用。第三章 评估从OL分离的细菌和真菌菌株的体外致癌特性。本研究将 由国际专家团队进行，将招募210名OL患者，105名OSCC患者和420名健康人。 在两个研究中心招募对照受试者（共735例受试者）。一千五百口水 并使用鸟枪宏基因组学和元转录组学分析来分析拭子样品。此外，委员会认为， 将筛选36株先前从具有不同程度异型增生的OL患者中分离的细菌/真菌菌株 对细胞增殖、侵袭、上皮-间质转化、细胞因子和基质的影响 体外正常和发育异常上皮细胞的金属蛋白酶产生。这项研究将确定 在口腔癌发生的早期阶段的微生物组，并确定OL和 在随后的研究中，其可以被验证为MT的生物标志物。

Project Summary There is increasing interest in the role of the microbiome in the development and/or progression of various cancers including oral squamous cell carcinoma (OSCC). Research has shown that OSCC is associated with a distinct microbiome. However, OSCC in itself represents a late stage in a disease process in which the observed microbial changes are as likely to be passenger or, at best, late driving events. To better understand the role of the microbiome in OSCC and identify microbial biomarkers for early detection and prevention, it is crucial to assess the microbiome in early stages of the disease, e.g. oral potentially malignant disorders (OPMDs). Oral leukoplakia (OL; the focus of this study) is the most common OPMD. Little is known about the microbiome associated with OPMDs, its role in their progression, or its potential as non-invasive diagnostic biomarker to complement oral epithelial dysplasia grading, currently the best available indicator. Our long-term goal is to identify diagnostic/prognostic microbial biomarkers of OPMDs and determine the effect of the microbiome on oral carcinogenesis by conducting a large-scale, international consortium study in two phases: a case-control study (this application) to comprehensively study the dysbiotic microbiome associated with OL/dysplasia in a North American population; followed by a longitudinal study (future application) that will involve following up the same and additional cohorts of OPMDs to identify/validate microbiome biomarkers of malignant transformation (MT). Our hypothesis is that shifts in microbiome composition and function associated with OL represent early driver events that correlate with the severity of dysplasia and contribute to the progression of oral cancer. In addition, we also hypothesize that oncogenic strains of oral microorganisms could be driving these disorders. To address these hypotheses, we propose 3 specific aims: 1) Characterize the composition of the multi-kingdom microbiome associated with OL and the different grades of dysplasia; 2) Define both microbial and host transcriptomes in OL and potential interactions between these two segments of the metatranscriptome. 3) Assess in vitro the oncogenic properties of bacterial and fungal strains isolated from OL. The study will be conducted by a team of international experts and will enroll 210 OL patients, 105 OSCC patients, and 420 healthy control subjects (735 subjects in total) to be recruited in two study centers. One-thousand and five hundred saliva and swab samples will be analyzed using shotgun metagenomic and metatranscriptome analysis. Furthermore, 36 bacterial/fungal strains previous isolated from OL patients with different grades of dysplasia will be screened for their effect on proliferation, invasion, epithelial-mesenchymal transition and cytokine and matrix metalloproteinases production by normal and dysplastic epithelial cells in vitro. The study will establish the role of the microbiome in the early stages of oral carcinogenesis, and identify microbial biomarkers of OL and dysplasia, which in a subsequent study can be validated as biomarkers of MT.