P. gingivalis genes essential for tobacco smoke survival

项目总结 牙周炎是一种由微生物驱动的牙齿周围组织的破坏性疾病，大约发生在 50%的人口，并与几种衰弱的全身疾病有关(包括血管和肺部疾病， 糖尿病和早产)。吸烟的患者患牙周炎的易感性增加，而且更有可能 比不吸烟的人表现出严重的疾病和难以治疗。事实上，最新的流行病学证据 这表明吸烟是发达国家破坏性牙周病病例的主要原因。吸烟会增加感染 增加牙周致病菌牙龈卟啉单胞菌的含量和数量。然而，潜在的机制 这一现象需要澄清。 我们计划为两个牙龈假单胞菌菌株(Atcc 33277)建立全基因组转座子测序文库 和W83)被用来(I)鉴定被认为是牙龈假单胞菌在香烟烟雾中存活所必需的基因 (Ii)通过产生单基因缺失突变体和 监测它们在香烟诱导的压力下在浮游植物和生物膜中生长的能力；(Iii)测定体内的 这些突变与烟雾加重的牙周炎小鼠模型的相关性；(Iv)建立必要的 多个低传代临床牙龈假单胞菌的基因；(V)确定这些基因的功能 在体外和体内对烟草诱导的压力的存活和(Vi)在硅胶中鉴定其他 其毒力在吸烟者身上也会增强的细菌。 成功的R01有几个潜在的翻译好处，包括更好地理解P。 牙周杆菌在富含烟草毒素的环境中茁壮成长；未来针对必要基因的治疗靶点以控制牙龈假单胞菌感染 在吸烟者中；预防或控制牙龈假单胞菌感染的新疫苗靶点的潜在鉴定；替代疗法 基于对吸烟引起的和/或加重的牙周疾病的机械性洞察力为吸烟者提供的养生法；以及 建立烟草强化病原菌基本基因数据库，为鉴定常见致病菌提供便利 在烟草烟雾暴露中存活的细菌策略，从而扩大了研究的意义，超出了口腔 空洞。

PROJECT SUMMARY Periodontitis, a microbial-driven, destructive disease of the tissues surrounding the teeth, occurs in approximately 50% of the population and is associated with several debilitating systemic conditions (including vascular and lung diseases, diabetes mellitus, and pre-term birth). Patients who smoke have increased susceptibility to periodontitis and are more likely than non-smokers to display severe disease and to be refractory to treatment. Indeed, the most recent epidemiological evidence suggests that tobacco smoking accounts for the majority of destructive periodontal disease cases in developed nations. Smoking enhances infection rates and enriches numbers of the keystone periodontal pathogen, Porphyromonas gingivalis. However, the mechanisms underlying this phenomenon are in need of elucidation. We plan to generate whole genome transposon sequencing (TnSeq) libraries for two P. gingivalis strains (ATCC 33277 and W83) to be employed in order to (i) identify genes that are putatively essential for P. gingivalis to survive cigarette smoke exposure in vitro; (ii) to validate that TnSeq-identified genes are indeed essential by generating single gene deletion mutants and monitoring their ability to grow planktonically and in biofilms under cigarette-induced stress; (iii) to determine the in vivo relevance of such mutations in a murine model of smoke-exacerbated periodontitis; (iv) to establish the distribution of essential genes in multiple low-passage clinical P. gingivalis isolates; (v) to characterize the function of those genes confirmed to be requisite for survival of tobacco-induced stress both in vitro and in vivo and (vi) to identify, in silico, essential gene orthologues in other bacteria whose virulence is also enhanced in tobacco smokers. There are several potential translational benefits to a successful R01, which include a better understanding of how P. gingivalis thrives in a tobacco-toxin rich environment; future therapeutic targeting of essential genes to control P. gingivalis infection in smokers; potential identification of novel vaccine targets for prevention or control of P. gingivalis infection; alternate treatment regimens for smokers based on mechanistic insight into smoke-induced and/or exacerbated periodontal diseases; and the establishment of an essential gene database for tobacco-enhanced pathogens that will facilitate the identification of common bacterial strategies for surviving tobacco smoke exposure, thus, broadening the significance of the research beyond the oral cavity.