Probing the architecture, assembly, and function of amyloid-polysaccharide entanglements in bacterial biofilms

探究细菌生物膜中淀粉样蛋白-多糖缠结的结构、组装和功能

基本信息

批准号：
10605820
负责人：
Schuyler A. Chambers
金额：
$ 6.91万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10605820
关键词：
Acceleration Address Adhesives Ammonium Amyloid Amyloid Proteins Amyloid fibers Anti-Bacterial Agents Anti-Infective Agents Antibiotics Architecture Arginine Bacteria Bacterial Polysaccharides Binding Biochemical Biological Assay Biophysics Biopolymers Cell surface Cells Cellulose Chemicals Communities Creativeness Detection Development Diffusion Disease Disinfectants Escherichia coli Evaluation Event Exhibits Extracellular Matrix Extracellular Protein Fellowship Fluorescence Microscopy Foundations Future Human Microbiome Infection Institution Investigation Isotope Labeling Link Magnetic Resonance Mechanics Mentorship Microbial Biofilms Modification Molecular NMR Spectroscopy Nature Nuclear Magnetic Resonance Organism Pathogenesis Pathology Polymers Polysaccharides Production Research Research Project Grants Role Salmonella Salmonella enterica Sampling Structure Symbiosis Testing Time Tissues Training Universities Urinary tract Vancomycin Visualization Western Blotting Work bacterial community cell community clinically relevant cohesion commensal bacteria design efficacy evaluation exhaust experimental study extracellular immunoregulation in vivo member microbial microbiome multidisciplinary mutant novel pathogen pathogenic bacteria phosphoethanolamine pressure recruit solid state nuclear magnetic resonance

项目摘要

Project Summary Bacteria are most commonly found in nature in multicellular communities termed biofilms. Biofilms are formed when bacteria synthesize, secrete, and enmesh themselves with diverse biopolymers. Beneficial bacteria in the microbiome assemble biofilms, while biofilms are unfortunately also linked to difficult-to-treat infections that exhibit increased tolerance to antibacterials and can exhaust treatment options. However, there are no blueprints for how bacteria build these tissue-like architectures and uncovering these details can accelerate discovery of new anti-infectives. E. coli, in particular, are normal residents in the healthy microbiome, but emerge as pathogens when they egress and colonize the urinary tract. E. coli, Salmonella species and other Gram-negative organisms harness specific amyloid and polysaccharide machinery to elaborate mechanically robust extracellular matrix architectures resembling baskets and blankets that surround cells and drive the formation of tissue-like biofilms. Due to the complexity of biopolymer composites, there are significant challenges associated with studying their structure and function, yet the ubiquity of these biopolymers makes them of high importance for study. This research plan is directed to test molecular hypotheses for how bacteria employ curli and phosphoethanolamine cellulose, a newly discovered chemically modified form of cellulose, to enmesh themselves in extracellular matrix (ECM). The research plan will test hypotheses regarding functional roles that we propose are ascribed to the zwitterionic phosphoethanolamine modification. Aim 1 is directed to evaluate the temporal and spatial developments of matrix assembly beyond the bacterial cell surface using fluorescence microscopy and creative functional biochemical assays. Aim 2 will implement a strategically designed solid-state nuclear magnetic resonance (NMR) approach to detect molecular contacts between polysaccharides and protein amyloids that are responsible for matrix cohesion. The functional benefit of ECM biopolymers will be determined in Aim 3, where clinically relevant antibiotics and a novel vancomycin-conjugate will be evaluated for efficacy against pEtN cellulose and curli containing biofilms. This work promises to formulate a molecular foundation for future avenues of inquiry at the host-pathogen interface, involving possible immunomodulatory roles of bacterial polysaccharides and amyloids, and possible biopolymer contributions to microbiome symbiosis and amyloid- associated disease pathologies. The fellowship candidate will receive significant training in solid-state NMR spectroscopy to study molecular interactions within E. coli biofilms and biochemical approaches to investigate bacterial communities. The considerable support and mentorship structure provided through this fellowship, the research sponsor (Prof. Lynette Cegelski) and institution (Stanford University) will facilitate the professional development of the fellowship applicant and the rigorous scientific investigation of the proposed research.

项目摘要细菌在自然界中最常见于称为生物膜的多细胞群落。形成生物膜当细菌合成时，分泌并与不同的生物聚合物融合在一起。有益细菌微生物组组装生物膜，而生物膜也与难以治疗的感染有关表现出对抗菌物质的耐受性，可以耗尽治疗选择。但是，没有蓝图有关细菌如何建立这些类似组织的结构并发现这些细节可以加速发现新的反感染者。尤其是大肠杆菌是健康微生物组中的正常居民，但出现为病原体在泌尿道出口和定居时。大肠杆菌，沙门氏菌和其他革兰氏阴性有机体线束特异性淀粉样蛋白和多糖机械机械稳定性细胞外基质体系结构类似于围绕细胞的篮子和毯子，并驱动形成组织样生物膜。由于生物聚合物复合材料的复杂性，因此存在重大挑战随着研究它们的结构和功能，这些生物聚合物的无处不在使它们非常重要进行研究。该研究计划用于测试细菌如何采用卷和的分子假设磷酸乙醇胺纤维素是一种新发现的化学改性形式的纤维素，以enmesh 自己在细胞外基质（ECM）中。研究计划将检验有关功能角色的假设我们建议归因于际离子磷酸乙醇胺的修饰。 AIM 1被指示评估使用荧光超出细菌细胞表面的基质组件的时间和空间发展显微镜和创意功能性生化测定。 AIM 2将实施战略设计的固态核磁共振（NMR）方法可检测多糖和蛋白质之间的分子接触负责基质内聚的淀粉样蛋白。将确定ECM生物聚合物的功能益处在AIM 3中，将评估与临床相关的抗生素和新型万古霉素 - 偶联物的功效针对含有生物膜的Petn纤维素和卷卷。这项工作有望为未来在宿主 - 病原体界面进行查询的途径，涉及细菌的免疫调节作用多糖和淀粉样蛋白，以及可能对微生物组共生和淀粉样蛋白的生物聚合物贡献相关的疾病病理。奖学金候选人将接受固态NMR的重大培训研究大肠杆菌生物膜内分子相互作用的光谱和生化方法细菌群落。通过此团契提供的大量支持和指导结构，研究赞助商（Lynette Cegelski教授）和机构（斯坦福大学）将促进专业人士奖学金申请人的发展和对拟议研究的严格科学研究。