P. gingivalis genes essential for tobacco smoke survival

牙龈卟啉单胞菌基因对烟草烟雾生存至关重要

• 批准号: 9331181
• 负责人: David A Scott
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9331181
• 关键词: Adult; Anaerobic Bacteria; Bacteria; Bacterial Model; Biochemical; Biological Assay; Chronic; Cigarette; Clinical; Collection; Communicable Diseases; Complex; Computer Simulation; Crystallography; Data; Databases; Deposition; Developed Countries; Developing Countries; Diabetes Mellitus; Disease; Dose; Economic Burden; Ensure; Environment; Epidemiology; Essential Genes; Exhibits; Exposure to; Future; Gene Deletion; Generations; Genes; Genome; Growth; Homologous Gene; Immune; In Vitro; Individual; Infection; Inflammation; Investigation; Laboratories; Libraries; Lung diseases; Microbial Biofilms; Modeling; Monitor; Mus; Mutation; New Zealand; Nicotine; Oral; Oral cavity; Output; Pathogenicity; Patients; Peptide Hydrolases; Periodontal Diseases; Periodontitis; Periodontium; Persons; Physiology; Population; Porphyromonas gingivalis; Predisposition; Premature Birth; Prevalence; Prevention; Preventive; Process; Refractory; Research; Resistance; Scandinavia; Smoke; Smoker; Smoking; Stress; Structural Models; Techniques; Testing; Time; Tissues; Tobacco; Tobacco smoke; Tobacco smoking; Tooth structure; Toxin; Treatment Protocols; Variant; Vascular Diseases; Virulence; Work; base; cigarette smoking; environmental tobacco smoke exposure; enzyme activity; fitness; in vivo; insight; microbial; mouse model; mutant; non-smoker; novel; novel vaccines; pathogen; periodontopathogen; prediction algorithm; screening; therapeutic target; whole genome

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.

项目摘要 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， 50%的人群，并与几种使人衰弱的全身性疾病（包括血管和肺部疾病， 糖尿病和早产）。吸烟的患者对牙周炎的易感性增加， 比非吸烟者更容易出现严重疾病，并且难以治疗。事实上，最新的流行病学证据 研究表明，在发达国家，吸烟是造成破坏性牙周病的主要原因。吸烟增加感染 率和丰富的关键牙周病原体，牙龈卟啉单胞菌的数量。然而， 这一现象需要澄清。 我们计划为两种牙龈卟啉单胞菌菌株（ATCC 33277）产生全基因组转座子测序（TnSeq）文库 和W83），以便（i）鉴定牙龈卟啉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来验证TnSeq鉴定的基因确实是必需的， 监测它们在膨润土诱导的应力下在生物膜中的无张力生长的能力;（iii）测定它们在体内的生长速率， 这些突变在吸烟加重牙周炎小鼠模型中的相关性;（iv）建立必需的 多个低传代临床牙龈卟啉单胞菌分离株中的基因;（v）表征那些被证实是必需的基因的功能 烟草诱导的应激的存活在体外和体内，以及（vi）通过计算机模拟鉴定其他烟草诱导的应激中的必需基因直向同源物。 在吸烟者中毒性也增强的细菌。 成功的R 01有几个潜在的转化益处，包括更好地理解P。 牙龈卟啉单胞菌在富含烟草毒素的环境中生长旺盛;控制牙龈卟啉单胞菌感染的必需基因的未来治疗靶向 在吸烟者中;预防或控制牙龈卟啉单胞菌感染的新型疫苗靶点的潜在鉴定;替代治疗 基于对吸烟诱发和/或加重的牙周疾病的机理性洞察的吸烟者方案;以及 建立烟草强化病原体的基本基因数据库， 细菌策略生存烟草烟雾暴露，因此，扩大了研究的意义超越口腔 腔