Development and application of auxin-inducible degradation in Candida pathogens

念珠菌病原体生长素诱导降解的开发和应用

基本信息

批准号：
10742370
负责人：
MARK C HALL
金额：
$ 23.14万
依托单位：
PURDUE 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：
2023-08-24 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10742370
关键词：
Acceleration Animal Model Animals Antibiotics Antifungal Agents Aspergillus Auxins Benign Biology C-terminal CRISPR/Cas technology Candida Candida albicans Candida auris Candida glabrata Candidiasis Chemicals Clinical Communities DNA cassette Dependence Development Disease Outbreaks Disseminated candidiasis Dose Drug Targeting Drug resistance Effectiveness Engineering Evaluation Evolution Fluorescence Frequencies Fungal Drug Resistance Fungi Model Future Gene Deletion Genes Health Hospitals Human Immune Individual Infection Kinetics Larva Life Malignant Neoplasms Methods Modeling Molecular Biology Molecular Target Multi-Drug Resistance Mus Mycoses N-terminal Pathogenesis Performance Pharmaceutical Preparations Phosphoric Monoester Hydrolases Physiological Predisposition Protac Proteins Protocols documentation Reagent Reporter Reporting Research Saccharomyces cerevisiae Severities System Technology Testing Therapeutic Time Validation Virulence Work World Health World Health Organization analog chimera drug climate change conditional mutant detection platform drug development drug discovery drug resistant pathogen emerging pathogen experimental study functional genomics gene product gene repression genomic tools global health histone acetyltransferase immunosuppressed in vivo inhibitor interest mouse model new therapeutic target novel pathogen pathogenic fungus priority pathogen promoter protein function resistance factors secondary infection severe COVID-19 simulation technology platform therapeutic development tool

项目摘要

PROJECT SUMMARY Opportunistic fungal infection of immune-compromised individuals is an escalating world health problem, recently highlighted in a report from the World Health Organization. Lethal outbreaks of multi drug-resistant Candida auris in hospitals and the rise of drug resistance in normally benign commensal fungal species like Candida glabrata highlight the severity of the problem. Even severe COVID-19 cases facilitate secondary infection by fungal pathogens like Aspergillus and Candida that can be lethal. Current treatment options for fungal infections are limited to a few antifungal drug classes that are becoming increasingly ineffective. There is a pressing need for new molecular targets for antifungal development to deal with drug-resistant pathogens. Our central objective is to establish auxin-inducible degradation (AID) technology in Candida pathogens to enable functional studies of virulence and drug resistance factors and as a tool to facilitate antifungal target validation in the early stages of antifungal drug discovery. AID provides rapid and specific depletion of target proteins of interest and has key advantages over other common methods for protein functional characterization. In Aim 1 we will engineer molecular biology reagents and strains, and establish protocols, to validate and implement a modified AID system in C. albicans, C. glabrata, and C. auris. Our novel system should be applicable in any strain, including clinical isolates, of Candida pathogen species. Validation experiments will use novel virulence and drug resistance factors identified in our labs. In Aim 2 we will combine AID technology in Candida species with two common animal infection models, Galleria mellonella (waxworm) larvae and immunosuppressed mice, to create systems for early target validation and in vivo simulation of drug effects on pathogenesis. These systems will also be applied to our novel candidate antifungal targets. AID is a powerful functional genomics tool that will enable new research opportunities in fungal pathogens. Reagents and protocols established during the project will be made available to the research community, and the work will establish a blueprint for expanding AID use to other diverse fungal pathogens. Overall, this technological platform will address the pressing need for identification and validation of viable new targets for antifungal therapeutic development.

项目概要免疫受损个体的机会性真菌感染是一个不断升级的世界健康问题，最近世界卫生组织的一份报告强调了这一点。多重耐药菌致命爆发医院中的耳念珠菌以及正常良性共生真菌物种（如光滑念珠菌凸显了问题的严重性。即使是严重的 COVID-19 病例也会促进继发性曲霉菌和念珠菌等真菌病原体感染可能致命。目前的治疗方案真菌感染仅限于几种越来越无效的抗真菌药物。那里迫切需要开发新的抗真菌分子靶点来应对耐药病原体。我们的中心目标是在念珠菌病原体中建立生长素诱导降解（AID）技术能够进行毒力和耐药因素的功能研究，并作为促进抗真菌靶点的工具抗真菌药物发现的早期阶段的验证。 AID 可快速、特异性地消除靶标感兴趣的蛋白质，与其他常见的蛋白质功能方法相比具有关键优势表征。在目标 1 中，我们将设计分子生物学试剂和菌株，并建立方案，以在白色念珠菌、光滑念珠菌和耳念珠菌中验证并实施改良的 AID 系统。我们的新颖系统应适用于念珠菌病原体物种的任何菌株，包括临床分离株。验证实验将使用我们实验室确定的新毒力和耐药因子。在目标 2 中，我们将结合具有两种常见动物感染模型的念珠菌属 AID 技术，大蜡虫（蜡虫）幼虫和免疫抑制小鼠，创建用于早期靶标验证和药物体内模拟的系统对发病机制的影响。这些系统也将应用于我们的新型候选抗真菌靶点。援助是一个强大的功能基因组学工具将为真菌病原体的研究提供新的机会。试剂项目期间制定的协议将提供给研究界，并且工作将建立一个蓝图，将援助的使用范围扩大到其他多种真菌病原体。总体而言，这项技术该平台将满足识别和验证可行的抗真菌新靶点的迫切需求治疗的发展。