Structure and Function of C. Difficile Toxins

艰难梭菌毒素的结构和功能

基本信息

批准号：
9913462
负责人：
Min Dong
金额：
$ 80.4万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-15 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9913462
关键词：
Accounting Address Affinity Basic Science Binding Biology Bontoxilysin CRISPR/Cas technology Cell model Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Clostridium difficile Clostridium difficile tcdA protein Colitis Collaborations Complement Complex Crystallization Custom Data Developed Countries Development Diarrhea Disease Docking Epithelial Epithelium Exhibits Exotoxins FZD1 gene FZD2 gene Fatty Acids Gastroenteritis Goals Heparitin Sulfate Human Infection Investigation Knowledge Length Lipoprotein (a)Low Density Lipoprotein Receptor Mass Spectrum Analysis Mediating Modeling Molecular Mus Mutagenesis Nonesterified Fatty Acids Organoids Pathogenesis Pathogenicity Pathway interactions Proteins Publishing Receptor Cell Research Role Series Site Structural Models Structure Therapeutic Intervention Toxic effect Toxin Tropism WNT Signaling Pathway Work alpha Toxin antibiotic-associated diarrhea crosslink design experimental study genome wide screen insight interdisciplinary approach member molecular targeted therapies multidisciplinary mutant novel novel therapeutic intervention receptor receptor binding research study small molecule targeted treatment therapeutic development tool translational study

项目摘要

Project Summary Clostridium difficile infection (CDI) is the most common cause of antibiotic-associated diarrhea and gastroenteritis-associated death among developed countries, and it is classified as one of the top three “urgent threats” by the US Centers for Disease Control and Prevention (CDC). The disease associated with CDI is mainly mediated by two homologous exotoxins, TcdA and TcdB. They target and disrupt the colonic epithelium, leading to diarrhea and colitis. Receptor-binding is a critical step in the toxin’s action and largely determine toxin tropisms, and these receptors are also molecular targets for therapeutic interventions. The goal of our proposal is to identify and characterize toxin receptors, establish a structural basis for toxin-receptor recognition, and harness this knowledge to gain a mechanistic understanding of the biology, pathogenesis, and therapy of TcdA and TcdB. We will take a multi-disciplinary approach, bringing together the expertise in toxin structure/function of Dr. Rongsheng Jin’s group with the expertise in toxin receptor identification/pathogenesis of Dr. Min Dong’s group. This proposed work is built on both our published work on identifying Frizzled (FZD) proteins as TcdB receptors, and extensive preliminary data on toxin-receptor interactions. We will focus on three aims: (1) elucidate the structural basis and pathogenic relevance of TcdB-FZD recognition;; (2) understand the structure of TcdB and the influence of FZD binding;; and (3) identify and characterize novel TcdA receptors through a CRISPR/Cas9 mediated genome-wide screens. These proposed studies will provide a comprehensive understanding of toxin-receptor interactions and how they contribute to the toxin biology and pathogenesis of TcdA and TcdB.

项目摘要艰难梭菌感染（CDI）是抗生素相关腹泻的最常见原因以及发达国家胃肠炎相关的死亡，并被归类为美国疾病控制与预防中心（CDC）的前三名“紧急威胁”。这与CDI相关的疾病主要由两种同源外毒素TCDA和TCDB介导。它们靶向并破坏结肠上皮，导致腹泻和结肠炎。受体结合是毒素作用的关键步骤，并在很大程度上决定了毒素的偏向主义，这些接收器也是治疗干预措施的分子靶标。我们建议的目的是确定并表征毒素受体，建立毒素受体识别的结构基础，然后利用这些知识对生物学，发病机理和 TCDA和TCDB的治疗。我们将采用多学科的方法，将 Rongsheng Jin博士的毒素结构/功能专业知识，具有毒素专业知识 Min Dong博士小组的接收者鉴定/发病机理。这项建议的工作建立在我们发表的有关将毛躁（FZD）蛋白识别为TCDB受体的工作和广泛的毒素受体相互作用的初步数据。我们将重点关注三个目标：（1）阐明 TCDB-FZD识别的结构巴西语和致病性相关性；（2）了解结构 TCDB和FZD结合的影响；（3）识别和表征新型TCDA 通过CRISPR/CAS9介导的全基因组屏幕的接收器。这些提出的研究将对毒素受体相互作用以及如何提供全面的理解有助于TCDA和TCDB的毒素生物学和发病机理。