Interactive Omics: Black raspberry metabolites and the oral microbiome in smokers

描述（由申请人提供）：口腔癌每年影响近30万美国人，由于其预后差和复发率高，是一个严重的健康问题。此外，世界范围内的烟草流行导致每年新诊断出数以千计的口腔癌病例。因此，针对因接触烟草而罹患口腔癌风险最高的人群的预防战略是一个高度优先事项。我们的最终目标是为高风险人群采用一种基于食物的癌症预防策略，使其具有良好的长期依从性和有效性。口腔是一个开放的微生物生态系统，它是700多种细菌的宿主，这些细菌形成了称为生物膜的健康相容群落。我们之前已经证明，这些生物膜在吸烟者体内迅速富集致病菌，导致早期的高炎症反应。此外，戒烟逆转了这种致病性细菌的重新定殖，这表明吸烟对口腔微生物群有直接影响，并可能通过干扰正常的宿主-细菌相互作用而增加口腔致癌过程的风险。因此，我们的中心假设是口腔细菌和富含植物化学物质的黑树莓食品之间存在重要的双向相互作用，最终导致炎症的衰减和疾病的改善。首先，我们将通过纵向临床研究设计和一种新的BRB传递系统（BRB花蜜），结合霰弹枪宏基因组测序和计算生物信息学提供的分辨率，确定黑树莓植物化学物质（BRB）对口腔生物膜内群落动态的影响。接下来，我们将研究当前吸烟者和从不吸烟者口腔细菌群落对brb代谢的影响。靶向HPLC-MS/MS分析将用于鉴定已知的酚类物质和代谢物，而非靶向UHPLC Q-TOF代谢组分析将用于鉴定新的化合物。最后，我们将通过纵向临床研究设计和一种新的高通量“双RNA-Seq”方法来评估BRB及其代谢物在逆转吸烟对口腔宿主-微生物相互作用的影响方面的功效，同时量化暴露于BRB花蜜前后口腔细菌和口腔粘膜转录组的变化。在这些特定目标的成功完成之后，我们将首次证明brb能够以一种支持口腔癌预防策略的方式减轻吸烟诱导的致病性代谢过程和基因表达模式。

DESCRIPTION (provided by applicant): Oral cancer affects nearly 300,000 Americans each year and is a serious health concern due to its poor prognosis and high recurrence rate. Moreover, the worldwide tobacco epidemic is leading to thousands of new cases of oral cancer being diagnosed each year. Targeting preventive strategies for those at highest risk for oral cancer due to tobacco exposure is thus a high priority. Our ultimate goal is to employ a food-based cancer preventive strategy for high-risk populations that allows excellent long-term adherence and efficacy. The oral cavity is an open microbial ecosystem that plays host to over 700 species of bacteria that form health- compatible communities called biofilms. We have previously demonstrated that these biofilms are rapidly enriched for pathogenic bacteria in smokers, resulting in an early hyper-inflammatory response. Furthermore, smoking cessation reverses this pathogenic bacterial recolonization, demonstrating that smoking has a direct effect on the oral microbiome and may increase the risk for oral carcinogenic processes by disturbing normal host-bacterial interactions. Therefore, our central hypothesis is that a critical bi-directional interaction exists between oral bacteria and phytochemical-rich black raspberry food products, which ultimately results in attenuation of inflammation and amelioration of disease. First, we will determine the effect of black raspberry phytochemicals (BRBs) on community dynamics within oral biofilms by combining a longitudinal clinical study design and a novel BRB delivery system (BRB nectar) with the resolution provided by shotgun metagenomic sequencing and computational bioinformatics. Next, we will examine the effect of oral bacterial communities on metabolism of BRBs in current and never smokers. Targeted HPLC-MS/MS analysis will be used to identify known phenolics and metabolites, while untargeted UHPLC Q-TOF metabolome analysis will identify novel compounds. Finally, we will evaluate the efficacy of BRBs and their metabolites in reversing the effect of smoking on oral host-microbial interactions by combining a longitudinal clinical study design with a novel, high-throughput "dual RNA-Seq" methodology to simultaneously quantify both oral bacterial and oral mucosal transcriptome changes before and after exposure to BRB nectar. Following the successful completion of these Specific Aims, we will demonstrate for the first time the ability of BRBs to mitigate pathogenic metabolic process and gene expression patterns induced by cigarette smoke in a manner that supports oral carcinogenesis prevention strategies.