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.

项目概要 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，大约发生在 占人口的 50%，与多种使人衰弱的全身性疾病有关（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可能性更大，并且更容易患牙周炎。 比不吸烟者表现出严重疾病并且难以治疗。事实上，最新的流行病学证据 表明吸烟是发达国家破坏性牙周病病例的大部分。吸烟会加剧感染 牙周病的关键病原体——牙龈卟啉单胞菌的数量增加了。然而，其背后的机制 这种现象需要加以阐明。 我们计划为两种牙龈卟啉单胞菌菌株（ATCC 33277）生成全基因组转座子测序（TnSeq）文库 和 W83) 用于 (i) 鉴定假定牙龈卟啉单胞菌在香烟烟雾中存活所必需的基因 体外暴露； (ii) 通过生成单基因缺失突变体来验证 TnSeq 鉴定的基因确实是必需的，并且 监测它们在香烟引起的压力下浮游和生物膜中生长的能力； (iii) 体内测定 此类突变在烟雾加剧的牙周炎小鼠模型中的相关性； (iv) 确定必需品的分配 多个低传代临床牙龈卟啉单胞菌分离株中的基因； (v) 表征那些被确认为必要的基因的功能 用于在体外和体内抵抗烟草引起的应激，以及 (vi) 在计算机中识别其他基因中的重要基因直向同源物 吸烟者体内毒力也会增强的细菌。 成功的 R01 有几个潜在的转化好处，其中包括更好地了解 P. 牙龈杆菌在富含烟草毒素的环境中繁衍生息；未来治疗靶向控制牙龈卟啉单胞菌感染的必需基因 吸烟者；预防或控制牙龈卟啉单胞菌感染的新疫苗靶标的潜在鉴定；替代治疗 基于对烟雾诱发和/或加剧的牙周病的机制洞察而制定的吸烟者治疗方案；和 建立烟草增强病原体的基本基因数据库，这将有助于识别常见的病原体 细菌在烟草烟雾暴露中幸存的策略，从而扩大了该研究的意义，超越了口腔 空腔。