Systematic in vitro and in vivo genetic analysis of C.albicans protein kinases

白色念珠菌蛋白激酶的系统体外和体内遗传分析

基本信息

批准号：
10308519
负责人：
Damian J Krysan
金额：
$ 19.19万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10308519
关键词：
Address Affect Age Antifungal Agents Bar Codes Biological Assay CRISPR/Cas technology Candida albicans Candidiasis Cell surface Cells Collection Communities Complex Development Disease Disseminated candidiasis Drug Targeting Environment Eye Gastrointestinal tract structure Genetic Genetic Transcription Goals Heart Human Hyphae In Vitro Infection Kidney Knowledge Libraries Life Liver Mediating Microbial Biofilms Modeling Mucous Membrane Neuraxis Organ Pathogenesis Pathway Analysis Pathway interactions Persons Phosphoproteins Physiological Physiology Process Protein Analysis Protein Dynamics Protein Kinase Research Resources Role Skin Spleen System Therapeutic Virulence Yeast Model System base body system design experimental study fitness genetic analysis genetic resource gut colonization human pathogen in vivo inorganic phosphate irritation microbiota mouse model mutant novel strategies novel therapeutic intervention novel therapeutics pathogen pathogenic fungus programs transcription factor urogenital tract

项目摘要

PROJECT SUMMARY Candida albicans is both a commensal component of the normal human microbial flora and an important human pathogen. Although the commensal niches occupied by C. albicans are primarily limited to the gastrointestinal and genitourinary tracts, candidal disease, in contrast, can involve a wide range of organ systems including kidney, spleen, liver, heart, eye, the central nervous system, and skin/mucosae. As such, C. albicans is a versatile pathogen that adapts to a wide range of physiologically diverse environments within the human host. Understanding the systems that facilitate this adaptation is central to developing detailed mechanistic models of C. albicans pathogenesis and could inform the identification of new therapeutic approaches to candidiasis. To date, C. albicans transcription factors (TF) are the best characterized components of this system, due in part to the availability of comprehensive sets of TF deletion mutants. In contrast, upstream regulators of C. albicans physiology such as protein kinases (PK) have not been systematically studied to the same extent. To address this knowledge gap, the long-term goal of our project is to systematically characterize the role of PKs in C. albicans pathobiology. As a first step toward this goal, we propose to: 1) generate and validate a barcoded set of homozygous and heterozygous C. albicans PK deletion mutants using a transient CRISPR/Cas9 strategy; 2) Systematically characterize the PK deletion mutants in two murine model of candidiasis and two virulence relevant in vitro processes: biofilm and hyphae formation; 3) Identify and characterize the network of PK-TF pairs that regulate biofilm and hyphae formation using genetic interaction analysis. Successful completion of these aims will provide a useful genetic resource for the research community, provide the first systematic functional characterization of C. albicans PKs in vivo and in vitro, and set the stage for subsequent in-depth analyses of the genetic networks regulated by C. albicans PKs.

项目概要白色念珠菌既是正常人类微生物菌群的共生成分，也是重要的微生物菌群。人类病原体。尽管白色念珠菌占据的共生生态位主要限于相比之下，胃肠道和泌尿生殖道念珠菌病可累及广泛的器官系统包括肾、脾、肝、心、眼、中枢神经系统和皮肤/粘膜。因此，C. 白色念珠菌是一种多才多艺的病原体，可以适应广泛的生理多样化环境。人类宿主。了解促进这种适应的系统对于制定详细的白色念珠菌发病机制的机制模型，可以为新治疗方法的识别提供信息念珠菌病的治疗方法。迄今为止，白色念珠菌转录因子 (TF) 是特征最明确的该系统的组成部分，部分归因于全面的 TF 缺失突变体集的可用性。在相比之下，白色念珠菌生理学的上游调节因子如蛋白激酶（PK）尚未被研究。系统地研究到同样的程度。为了弥补这一知识差距，我们项目的长期目标是系统地表征 PK 在白色念珠菌病理学中的作用。作为实现这一目标的第一步，我们建议：1) 生成并验证一组带有条形码的纯合子和杂合子白色念珠菌 PK 删除使用瞬时 CRISPR/Cas9 策略的突变体； 2) 系统地表征 PK 缺失突变体两种念珠菌病鼠模型和两种毒力相关的体外过程：生物膜和菌丝形成； 3）使用遗传技术识别和表征调节生物膜和菌丝形成的 PK-TF 对网络交互分析。这些目标的成功完成将为人类提供有用的遗传资源。研究社区，提供了白色念珠菌 PK 体内和体内的第一个系统功能表征体外，并为后续深入分析白色念珠菌 PK 调节的遗传网络奠定了基础。