Complex haploinsufficiency based genetic analysis of C. albicans pathogenesis

基于复杂单倍体不足的白色念珠菌发病机制的遗传分析

基本信息

批准号：
10300444
负责人：
Damian J Krysan
金额：
$ 41.51万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10300444
关键词：
ACE2 AIDS/HIV problem Adhesions Affect Azole resistance BCR gene Bar Codes Biological Assay CD4 Lymphocyte Count Candida albicans Candidiasis Cells Complex Consensus Cyclic AMP-Dependent Protein Kinases Development Disease Epithelial Esophagus Filament Gene Expression Profile Genes Genetic Genetic Screening Genetic Transcription Genetic study Goals HIV Hyphae Immunosuppression In Vitro Infection Inflammation Libraries Mediating Mediator of activation protein Microbial Biofilms Molecular Morphogenesis Mouth Diseases Mucous Membrane Mus Mutation Oral candidiasis Pathogenesis Pathway Analysis Pathway interactions Patients Persons Pharmaceutical Preparations Phenotype Phosphorylation Site Phosphotransferases Plasmids Process Public Health Regulation Resources Signal Pathway Surface Testing Tissues Transcription Process Virulence Yeasts antiretroviral therapy base clinically significant compliance behavior deletion library gene function genetic analysis in vivo intravital imaging mouse model mutant novel novel therapeutic intervention opportunistic pathogen oropharyngeal thrush pathogenic fungus screening transcription factor

项目摘要

PROJECT SUMMARY Candida albicans is an important opportunistic pathogen for people living with HIV/AIDS and primarily causes mucosal diseases such as oropharyngeal candidiasis (OPC) and esophageal candidiasis (EC). In the era of antiretroviral therapy (ART), C. albicans remains the most common fungal pathogen affecting those with HIV/AIDS and OPC is one of the most common infections in HIV patients with mild-to-moderate immunosuppression (CD4 counts 200-500). The pathogenesis of OPC can be separated into two stages: 1) C. albicans adhesion/biofilm formation on the mucosal surface and 2) hyphae-mediated invasion of the epithelium and submucosal stroma with concomitant inflammation and tissue damage. Thus, OPC pathogenesis is dependent on three of the most important mediators of C. albicans virulence: adhesion, biofilm formation, and filamentation. Although each of these factors has been studied using specific C. albicans mutants, no systematic large-scale genetic analysis of OPC pathogenesis has been undertaken. The goal of this application is to define and characterize the transcriptional networks that underlay the ability of C. albicans to cause oral disease. Recently, we have pioneered the use of complex haploinsufficiency (CHI)-based genetic interaction analysis to understand the function of complex genetic networks. Here, we propose to use CHI analysis to test the hypothesis that Cbk1 represents a master regulator of transcriptional processes critical for OPC pathogenesis (Aim 1). In addition to this intra-pathway analysis, we will perform the first large-scale genetic screens for, and CHI analyses of, TF networks required for murine oral candidiasis (Aim 2) and in vivo filamentation (Aim 3). The screen in Aim 3 will utilize our novel intra-vital imaging assay to quantitatively differentiate between yeast and filamentous cells in the sub-epithelial stroma of mice. This screen will not only be the first in vivo C. albicans filamentation screen but also will allow us to distinguish TFs required for the invasion stage of OPC from TFs required for the adhesion/biofilm formation stage.

项目摘要白色念珠菌是艾滋病毒/艾滋病患者的重要机会性病原体，主要是原因粘膜疾病，例如口咽念珠菌病（OPC）和食管念珠菌病（EC）。在时代抗逆转录病毒疗法（ART），白色念珠菌仍然是影响患有患者的最常见的真菌病原体艾滋病毒/艾滋病和OPC是艾滋病毒中最常见的感染之一免疫抑制（CD4计数200-500）。 OPC的发病机理可以分为两个阶段：1）白色念珠菌粘附/生物膜在粘膜表面形成，2）菌丝介导的侵袭上皮和粘膜粘膜层，带有炎症和组织损伤。因此，OPC 发病机理取决于白色念珠菌毒力的三个最重要的介质：粘附，生物膜形成和细丝。尽管已经使用特定白色念珠菌研究了这些因素突变体，未进行系统的OPC发病机理的系统大规模遗传分析。目标该应用程序是为了定义和表征具有白色念珠菌能力的转录网络引起口腔疾病。最近，我们率先使用了基于复杂的单倍弥补（CHI）的遗传相互作用分析以了解复杂遗传网络的功能。在这里，我们建议使用chi 分析以检验CBK1代表转录过程的主要调节剂的假设至关重要 OPC发病机理（AIM 1）。除了这项内部轨道分析外，我们还将执行第一个大规模鼠口服念珠菌所需的TF网络的遗传筛选和CHI分析（AIM 2）和体内细丝（目标3）。 AIM 3中的屏幕将利用我们的新颖重要内成像测定法来定量区分小鼠的酵母和丝状细胞。这个屏幕不仅会成为第一个体内C.白色念珠菌细丝屏幕，但也将使我们能够区分TFS 粘附/生物膜形成阶段所需的TFS的OPC侵袭阶段。