Characterization of viridans group streptococci from endophthalmitis

眼内炎草绿色链球菌的特征

• 批准号: 10057004
• 负责人: MARY E MARQUART
• 金额: $ 19.38万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057004
• 关键词: Amino Acid Sequence; Antibiotics; Bacteria; Bacterial Proteins; Bacteriology; Bioinformatics; Case Study; Clinical; Collection; Deletion Mutagenesis; Development; Disease; Endophthalmitis; Enzymes; Erythrocytes; Etiology; Eye; Eye Infections; Future; Genbank; Gene Expression; Genes; Genetic; Genome; Genomics; Genus staphylococcus; Growth; Health; Heterogeneity; Horizontal Gene Transfer; Human; In Vitro; Infection; Injections; Investigation; Kinetics; Knowledge; Lead; Literature; Mining; Oligonucleotide Primers; Operative Surgical Procedures; Oral cavity; Organism; Oryctolagus cuniculus; Outcome; Pathogenesis; Patients; Peptide Hydrolases; Process; Proteins; Recombinant Proteins; Recombinants; Research; Resistance; Role; Sequence Analysis; Sheep; Streptococcus; Streptococcus Viridans Group; Streptococcus anginosus; Streptococcus gordonii; Streptococcus mitis; Streptococcus mutans; Streptococcus oralis; Streptococcus sanguis; Structure of retinal pigment epithelium; Substrate Specificity; Testing; Translating; Trauma; Variant; Virulence; Virulence Factors; Virulent; Visual; Vitreous humor; bacterial endophthalmitis; comparative; cytokine; epidemiology study; experimental study; extracellular; genome analysis; genome sequencing; in vivo; inhibitor/antagonist; normal microbiota; oral commensal; oral pathogen; outcome forecast; pan-genome; protein degradation; protein purification; therapeutic development; therapy development; transcriptome; transcriptome sequencing; transcriptomics; whole genome

Project Summary/Abstract The viridans streptococci are classically considered oral commensals with cariogenic potential; however, these bacteria are also listed among the most common species that cause bacterial endophthalmitis. Exogenous bacterial endophthalmitis can occur following trauma, ocular surgery, or ocular injections. Antibiotics are used to treat this disease, however, antibiotics are only able to kill the bacteria and do not ameliorate the damage to the eye. Streptococcal species in particular are associated with very poor visual outcomes; despite treatment, evisceration or enucleation commonly occur. The immediate objective of this application is to characterize endophthalmitis strains of the viridans streptococci and identify their virulence factors. This objective is intended to initiate study into the broader, long-term objectives of determining the mechanisms of pathogenesis of this important group of species and identify potential targets for future therapies. Two major aims are proposed: 1) to investigate the role of bacterial protease HtrA in endophthalmitis and determine its vitreous and bacterial protein substrates, and 2) to analyze the pangenome and virulence transcriptome of endophthalmitis- causing strains. The first aim will be tested by function analysis of recombinant HtrA variants cloned from the viridans strains and by deletion mutagenesis of htrA from htrA-positive strains, as well as genetic addition of htrA to htrA-negative strains. The second aim will employ whole genome sequencing of 21 strains followed by bioinformatics pangenome analysis to identify conserved genes across the collection, and RNA-seq in vivo and in vitro to determine the virulence transcriptome of this group of species. Accomplishment of these aims will springboard the identification of key factors involved in ocular infection by these species. This research is expected to benefit patient ocular health in the long-term because the knowledge to be gained can aid future therapeutic development.

项目概要/摘要 草绿色链球菌通常被认为是具有致龋潜力的口腔共生菌。然而，这些 细菌也被列为引起细菌性眼内炎的最常见物种之一。外源性 细菌性眼内炎可能发生在外伤、眼部手术或眼部注射后。使用抗生素 然而，为了治疗这种疾病，抗生素只能杀死细菌，并不能改善对身体的损害。 眼睛。特别是链球菌物种与非常差的视力结果有关；尽管接受治疗， 经常发生内脏或去核。该应用程序的直接目标是表征 草绿色链球菌眼内炎菌株并鉴定其毒力因子。这个目标是 旨在启动研究以确定发病机制的更广泛、长期的目标 对这一重要物种进行研究，并确定未来治疗的潜在目标。两个主要目标是 提出：1）研究细菌蛋白酶HtrA在眼内炎中的作用并确定其玻璃体和 细菌蛋白底物，2) 分析眼内炎的全基因组和毒力转录组- 造成紧张。第一个目标将通过从克隆的重组 HtrA 变体的功能分析来测试 草绿色菌株和通过 htrA 阳性菌株中 htrA 的缺失诱变，以及基因添加 htrA 至 htrA 阴性菌株。第二个目标是对 21 个菌株进行全基因组测序，然后 生物信息学泛基因组分析，以识别整个集合中的保守基因，以及体内和体内的RNA-seq 体外测定这组物种的毒力转录组。这些目标的实现将 确定与这些物种眼部感染有关的关键因素。这项研究是 预计将长期有益于患者的眼部健康，因为获得的知识可以帮助未来 治疗的发展。