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

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

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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.

项目摘要白色念珠菌既是正常人类微生物菌群的共同成分，又是重要的人病原体。尽管白色念珠菌所占据的共生壁细分市场主要限于相比之下包括肾脏，脾，肝脏，心脏，眼睛，中枢神经系统和皮肤/粘膜在内的系统。因此，C。白色唱片是一种多功能的病原体，可适应多种生理上不同的环境人类主持人。了解促进这种适应的系统对于开发详细的系统至关重要白色念珠菌发病机理的机械模型，可以告知新治疗的鉴定念珠菌病的方法。迄今为止，白色念珠菌的转录因子（TF）是最好的特征该系统的组成部分，部分是由于全面的TF缺失突变体的可用性。在对比，白色念珠菌生理学的上游调节剂，例如蛋白激酶（PK）系统地研究了同一程度。为了解决这个知识差距，我们项目的长期目标是系统地表征PK在白色念珠菌病理生物学中的作用。作为实现这一目标的第一步，我们提议：1）生成和验证一组纯合子和杂合子白色念珠菌PK缺失使用瞬态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的鼠模型和两个在体外过程中相关的毒力：生物膜和菌丝形成； 3）识别和表征使用遗传来调节生物膜和菌丝形成的PK-TF对网络相互作用分析。这些目标的成功完成将为研究界，提供了体内白色念珠菌PK的第一个系统功能表征体外，为随后的白色念珠菌PKS调节的遗传网络进行了深入的分析奠定了基础。