Outer Membrane Proteins of Pathogenic Oral Treponemes Inhibit Actin Rearrangement and Antimicrobial Functions of Neutrophils

致病性口腔密螺旋体外膜蛋白抑制中性粒细胞肌动蛋白重排和抗菌功能

• 批准号: 10616559
• 负责人: Natalie Anselmi
• 金额: $ 3.35万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10616559
• 关键词: Actins; Address; Adult; Affect; Air; Animal Model; Bacteria; Bacterial Proteins; Biological Assay; Cell Count; Cell physiology; Cells; Chemotaxis; Clinical; Co-Immunoprecipitations; Complex; Confocal Microscopy; Cytoplasmic Granules; Cytoprotection; Cytoskeleton; Development; Disease Progression; EEF1A2 gene; Elements; Engineering; Ensure; Environment; Enzyme-Linked Immunosorbent Assay; Equilibrium; Escherichia coli; Flow Cytometry; Goals; Health; Histology; Immune; Immune response; Immune system; Immunologic Techniques; Immunoprecipitation; Impairment; Inflammation; Inflammatory; Interdisciplinary Study; Irrigation; Knowledge; Lipids; Liquid substance; Measures; Membrane; Membrane Proteins; Mentors; Methods; Microbiological Techniques; Microfilaments; Microscopy; Modeling; Molecular Biology; Mus; Neutrophil Infiltration; Oral; Oral health; Order Spirochaetales; PIK3CG gene; PTEN gene; Pathogenicity; Periodontal Diseases; Periodontium; Phagocytosis; Phosphatidylinositols; Phosphoric Monoester Hydrolases; Property; Protein Isoforms; Proteins; Recombinants; Regulation; Research; Role; Sequence Homology; Signal Pathway; Signal Transduction; Surface; Testing; Tissues; Tooth Loss; Training; Treponema; Treponema denticola; United States; Universities; Virulence Factors; Wasps; Western Blotting; Work; antimicrobial; career; cohort; dysbiosis; improved; in vivo; insight; leucyl-phenylalanine; monomer; mutant; neutrophil; novel therapeutics; oral biofilm; oral commensal; oral spirochetes; oral tissue; pathogenic bacteria; polymicrobial biofilm; prevent; recruit; response; saliva sample; skill acquisition; therapeutically effective

Periodontal disease is a bacterially induced inflammatory condition affecting 47% of adults in the United States and is the number one cause of tooth loss worldwide. This condition is characterized by the destruction of tooth-supporting structures resulting from dysbiosis between the host immune system and the normally commensal oral biofilm. Treponema denticola, Treponema maltophilium, and Treponema lecithinolyticum are three understudied bacterial species abundant in the polymicrobial biofilm associated with severe periodontal disease, and a complete understanding of their pathogenic properties is lacking. T. denticola, the most-well- studied oral spirochete, has a prominent virulence factor: the major outer sheath protein (Msp) that dysregulates the functions of host cells, including neutrophils. T. maltophilium and T. lecithinolyticum have Msp-like outer membrane proteins called MspA and MspTL, respectively. Neutrophils are key innate immune cells that protect oral tissues from pathogenic bacteria by coordinating cellular signaling, structural elements, and cell function. Msp inhibits neutrophil function by disrupting the balance of phosphoinositides, cellular lipid metabolites key for intracellular signaling. This disruption involves altered regulation of the PI3 kinase (PI3K) and phosphatase and tensin homolog (PTEN) axis, leading to inappropriate remodeling of the actin cytoskeleton, impaired chemotaxis, and altered functioning of neutrophils. A remaining gap in our knowledge is how these understudied Treponema proteins modulate actin dynamics to impair other crucial neutrophil properties. The overall objective of this project is to characterize how these Treponema species and their surface proteins manipulate neutrophil cytoskeleton signaling pathways and granule release to promote survival. We hypothesize that Msp-like proteins dysregulate actin remodeling in neutrophils to promote bacterial survival. To test this hypothesis, this project aims to (1) characterize the effects of Msp proteins on the PI3K/PTEN axis and actin branching dynamics and (2) assess the ability of Treponema species and their Msp proteins to promote survival by modulating neutrophil recruitment and degranulation. To achieve these aims, I will utilize a variety of methods, including analyses of actin incorporation, immunological techniques, microscopy, flow cytometry, animal models, molecular biology, and microbiological techniques. Completion of this project will provide valuable insight into the interactions between spirochetes and the immune system and how these relationships drive disease progression. The mentoring and training plan to be performed within the multidisciplinary research environment at the University at Buffalo will provide me with the scientific and professional development skills necessary to successfully transition to the next stage of a successful research career.

牙周病是一种细菌引起的炎症，影响美国 47% 的成年人 是全球牙齿脱落的第一大原因。这种情况的特点是破坏 由于宿主免疫系统与正常免疫系统之间的生态失调而产生的牙齿支持结构 共生口腔生物膜。齿状密螺旋体、嗜麦芽密螺旋体和解卵磷脂密螺旋体是 与严重牙周病相关的多种微生物生物膜中富含三种正在研究的细菌种类 疾病，并且缺乏对其致病特性的完整了解。 T. denticola，最好的- 研究口腔螺旋体，具有显着的毒力因子：主要外鞘蛋白（Msp） 失调宿主细胞的功能，包括中性粒细胞。 T. maltophilium 和 T. lecithinolyticum 有 Msp 样外膜蛋白分别称为 MspA 和 MspTL。中性粒细胞是关键的先天免疫 通过协调细胞信号、结构元件、保护口腔组织免受病原菌侵害的细胞 和细胞功能。 Msp 通过破坏磷酸肌醇、细胞脂质的平衡来抑制中性粒细胞功能 代谢物是细胞内信号传导的关键。这种破坏涉及 PI3 激酶 (PI3K) 调节的改变 磷酸酶和张力蛋白同源物 (PTEN) 轴，导致肌动蛋白的不适当重塑 细胞骨架、趋化性受损和中性粒细胞功能改变。我们的知识中尚存的空白是 这些正在研究的密螺旋体蛋白如何调节肌动蛋白动力学以损害其他重要的中性粒细胞 特性。该项目的总体目标是描述这些密螺旋体物种及其 表面蛋白操纵中性粒细胞骨架信号通路和颗粒释放以促进 生存。我们假设 Msp 样蛋白失调中性粒细胞中的肌动蛋白重塑，从而促进细菌 生存。为了检验这一假设，该项目旨在 (1) 表征 Msp 蛋白对 PI3K/PTEN 轴和肌动蛋白分支动力学和 (2) 评估密螺旋体物种及其 Msp 的能力 蛋白质通过调节中性粒细胞募集和脱粒来促进存活。为了实现这些目标，我 将利用多种方法，包括肌动蛋白掺入分析、免疫学技术、 显微镜、流式细胞术、动物模型、分子生物学和微生物技术。完成 该项目将为螺旋体和免疫系统之间的相互作用提供有价值的见解 这些关系如何驱动疾病进展。指导和培训计划将在 布法罗大学的多学科研究环境将为我提供科学和 成功过渡到成功研究的下一阶段所需的专业发展技能 职业。