Complex haploinsufficiency based genetic analysis of C. albicans pathogenesis

项目总结 白色念珠菌是艾滋病毒/艾滋病患者的一种重要的机会性病原体，主要原因是 粘膜疾病，如口咽部念珠菌病(OPC)和食道念珠菌病(EC)。在……时代 在抗逆转录病毒疗法(ART)中，白色念珠菌仍然是影响患者的最常见真菌病原体 HIV/AIDS和OPC是感染HIV患者中最常见的轻到中度感染之一 免疫抑制(CD4计数为200-500)。OPC的发病机制可分为两个阶段：1) 白色念珠菌在粘膜表面的黏附/生物膜形成和2)菌丝介导的侵袭 上皮和粘膜下间质伴有炎症和组织损伤。因此，OPC 致病机制依赖于白念珠菌毒力的三个最重要的媒介：黏附、生物被膜 形成和成丝。尽管这些因素中的每一个都已经用特定的白色念珠菌进行了研究 目前，尚未对OPC的发病机制进行系统的大规模遗传分析。的目标是 这项应用是为了定义和表征支持白念珠菌能力的转录网络。 导致口腔疾病。最近，我们率先使用了基于复杂单倍体不足(CHI)的基因 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 对Cbk1代表转录过程的主要调控因子这一假设进行分析以检验这一假设 OPC发病机制(目标1)。除了这种路径内分析之外，我们还将执行第一个大规模 小鼠口腔念珠菌病所需的转铁蛋白网络的遗传筛选和CHI分析(AIM 2)和体内 丝状化(目标3)。Aim 3中的筛查将利用我们新的活体成像分析来定量 分化小鼠上皮下基质中的酵母细胞和丝状细胞。此屏幕不仅将 作为体内第一个白念珠菌丝状筛选，也将使我们能够区分 黏附/生物膜形成阶段所需的因子对OPC的侵袭阶段。

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.