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: 10437631
• 负责人: David A Scott
• 金额: $ 36.8万
• 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 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10437631
• 关键词: Abscess; Adult; Alveolar Bone Loss; Anaerobic Bacteria; Award; Bacteria; Basic Science; Biological Assay; Biological Process; Blood Vessels; Cells; Chronic; Cigarette; Clinical; Communicable Diseases; Complex; Consumption; Data; Databases; Developed Countries; Diabetes Mellitus; Disease; Dose; Economic Burden; Environment; Epidemiology; Epithelial; Epithelial Cells; Essential Genes; Exhibits; Exposure to; Financial cost; Funding Mechanisms; Future; Gene Deletion; Genes; Genome; Gingiva; Growth; Health; Homologous Gene; Immune; Immune response; In Vitro; Infection; Inflammation; Libraries; Lung diseases; Modeling; Mouth Diseases; Mus; National Institute of General Medical Sciences; New Zealand; Nicotine; Oral; Oral cavity; Patients; Peptide Hydrolases; Periodontal Diseases; Periodontitis; Persons; Physiology; Population; Porphyromonas gingivalis; Predisposition; Premature Birth; Prevention; Refractory; Research; Resistance; Resources; Scandinavia; Smoke; Smoker; Smoking; Standardization; Stress; Subcutaneous abscess; System; Testing; Tissues; Tobacco; Tobacco smoke; Tobacco smoking behavior; Tooth structure; Toxin; Treatment Protocols; Variant; Vascular Diseases; Virulence; Work; base; biological adaptation to stress; cigarette smoke; cigarette smoke-induced; fitness; immune resistance; in vivo; infection rate; insight; mouse model; mutant; non-smoker; novel; novel vaccines; oral cavity epithelium; pathogen; periodontopathogen; success; therapeutic target; tobacco regulation; tobacco smoke exposure; trait; transcriptome; transcriptome sequencing; translational potential; transposon sequencing

PROJECT SUMMARY/ABSTRACT Periodontitis is a highly prevalent disease associated with several debilitating systemic conditions, including vascular and lung diseases, diabetes mellitus, and pre-term birth. 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 a key periodontal pathogen, Porphyromonas gingivalis. However, the mechanisms underlying this phenomenon are in need of elucidation. In the first iteration of award, facilitated by a two year NIGMS-funding mechanism, we generated an 80,000 colony transposon sequencing library for P. gingivalis strains ATCC 33277 and exploited this resource to: (i) Successfully identify 256 genes that are putatively essential for surviving tobacco-induced stress; (ii) Delineate a “core stress genome” commonly required for tobacco-survival, epithelial colonization and subcutaneous abscess formation; and (iii) Validated several genes as conditionally essential for P. gingivalis to survival of cigarette smoke- induced stress in vitro by generating single gene deletion mutants that exhibit either reduced absolute fitness in competition assays with the parental P. gingivalis strain in smoked medium; (iv) We have also generated a novel murine model of tobacco-exacerbated periodontitis. We now plan to build on these successes via three specific aims, as follow: Aim 1: Further elucidate P. gingival CSE-related survival strategies by generating mutants in single genes that are both CSE-essential (TnSeq data) ;and tobacco-regulated (RNAseq and qPCR); yet not compromised in their ability to colonize oral epithelial cells (TIGK model) or immune resistance (murine abscess model). Aim 2: Determine in vivo relevance of putatively CSE-essential genes using a murine smoke-exposure chamber model in a chronic Baker model of P. gingivalis-induced periodontitis. Aim 3: Establish the broad applicability of data gathered in Aims 1 and 2 by confirming biological function and distribution of CSE essential genes in multiple P. gingivalis strains. Long term advances are likely to include (a) A better understanding of how P. gingivalis thrives in a tobacco-toxin rich environment; (b) Future therapeutic targeting of essential genes to control P. gingivalis infection in smokers (CSE-essentiality data) and in general (multiple disease-relevant conditions, core stress genome data); (c) Alternate treatment regimens for smokers based on mechanistic insight into smoke-induced and/or exacerbated periodontal diseases;and (d) 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/摘要 牙周炎是一种高度普遍的疾病，与几种衰弱的全身状况有关， 包括血管疾病和肺部疾病，糖尿病和早产。最近的流行病学 有证据表明，烟草吸烟占大多数破坏性牙周疾病病例 发达国家。吸烟增强感染率并丰富了关键牙周病原体的数量， 牙龈卟啉单胞菌。但是，这种现象的基础机制需要阐明。在 由两年的Nigms资助机制编写的第一次裁决，我们产生了80,000个殖民地 用于牙龈疟原虫菌株ATCC的转座子测序库33277，并将此资源利用为： （i）成功识别256个基因，这些基因对于烟草引起的应激至关重要； （ii）描绘烟草育种，上皮定殖所需的“核心应力基因组” 和皮下脓肿的形成；和 （iii）验证了几个基因是牙龈疟原虫在香烟烟雾中存活至关重要的 通过产生单个基因缺失突变体，诱导应激在体外，该突变体暴露于降低 在烟培养基中与父母牙龈疟原虫菌株的竞争测定中的绝对适应性； （iv）我们还产生了一种新型的烟草牙周炎的鼠模型。 现在，我们计划通过以下三个特定目标来建立这些成功，如下： AIM 1：通过在单个基因中产生突变体，进一步阐明了牙龈P.牙龈P. 两者都是CSE- ENDERAINS（TNSEQ数据）和烟草调节的（RNASEQ和QPCR）；但未妥协 它们使口服上皮细胞（TIGK模型）或免疫耐药性（鼠脓肿模型）的能力。 AIM 2：使用鼠烟雾暴露确定推定的CSE代表基因的体内相关性 牙龈疟原虫诱导的牙周炎的慢性面包师模型中的腔室模型。 目标3：通过确认生物学功能，确定目标1和2中收集的数据的广泛适用性 CSE基因在多个牙龈疟原虫菌株中的分布。 长期进步可能包括（a）更好地理解牙龈牙龈疟原虫在A中如何壮成长 烟草蛋白丰富的环境； （b）将来对牙龈疟原虫的基本基因的未来治疗靶向 吸烟者的感染（CSE-肯定数据）和一般（多种疾病条件，核心应力） 基因组数据）； （c）基于对烟雾引起的机械洞察力的吸烟者的替代治疗方案 和/或恶化的牙周疾病；（d）建立用于烟草的基本基因数据库 增强的病原体将准备鉴定生存烟草的常见细菌策略 因此，烟雾暴露范围扩大了研究的重要性。