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

• 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
• 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

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.

项目摘要 免疫受损个体的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中强调了这一点。致命的耐多药病毒爆发 医院中的耳念珠菌和正常良性真菌种类如 光滑念珠菌突出了问题的严重性。即使是严重的COVID-19病例也会导致继发性 真菌病原体如曲霉菌和念珠菌感染，可能是致命的。的当前治疗选择 真菌感染仅限于几类抗真菌药物，它们变得越来越无效。那里 是一个迫切需要新的分子靶点的抗真菌发展，以应对耐药病原体。 我们的中心目标是建立念珠菌属病原体中的生长素诱导降解（AID）技术， 使毒力和耐药因子的功能研究成为可能，并作为促进抗真菌靶向的工具 在抗真菌药物发现的早期阶段进行验证。AID提供快速和特异性的靶细胞耗竭 目的蛋白质，并具有优于其他常见的蛋白质功能方法的关键优势 特征化在目标1中，我们将设计分子生物学试剂和菌株，并建立方案， 用C语言验证并实现了一个改进的AID系统。白色念珠菌、C.光叶C.耳。我们的新系统 应适用于念珠菌属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鉴定的新型毒力和耐药性因子。在目标2中，我们将联合收割机 AID技术在念珠菌属中的应用两种常见的动物感染模型，大蜡螟（蜡虫） 幼虫和免疫抑制小鼠，以创建早期靶点验证和药物体内模拟系统 对发病机制的影响。这些系统也将应用于我们的新的候选抗真菌靶点。援助是一项 强大的功能基因组学工具，将使新的研究机会，在真菌病原体。试剂 在项目期间建立的协议将提供给研究界， 将建立一个蓝图，扩大艾滋病的使用，以其他不同的真菌病原体。总的来说，这项技术 该平台将满足识别和验证抗真菌药物可行新靶点的迫切需求。 治疗发展