CANDIDA GENE EXPRESSION DURING OROPHARYNGEAL INFECTION

描述(申请人提供)：口咽念珠菌病(OPC)是艾滋病的首批临床表现之一。在发展为艾滋病的过程中，它发生在50-95%的HIV+个体中，据疾病预防控制中心估计，艾滋病患者的总体流行率为9%-31%。这种感染主要是由白色念珠菌引起的，这是一种共生真菌，可导致粘膜和侵袭性感染。我们的总体目标是了解控制宿主相互作用并参与OPC发病机制的白色念珠菌基因产物。我们将使用一种新的基因表达分析技术，纳米串核糖核酸测量，来确定在广泛使用的小鼠OPC模型中感染期间白色念珠菌的基因表达水平。我们的初步结果与以前的患者样本和体外感染基因表达有很好的相关性 数据，但也揭示了广泛的应激反应基因表达和转录因子-靶基因关系的重新连接。建议的研究集中在两类基因产物：转录因子和表面蛋白和分泌蛋白。我们之所以选择转录因子基因，是因为它们是一个高度调控的基因集，它们的产物已经或能够通过系统的突变筛选、表达谱和染色质免疫沉淀与功能相连。我们之所以选择表面和分泌蛋白基因，是因为它们的产物与宿主细胞直接相互作用，也因为它们提供了最容易获得的治疗靶点。我们将开发一个探索这些基因表达的参考数据集，并通过功能验证扩展分析。我们的发现将产生新的假设，并为社区中所有研究人员可以追寻的基因和途径的选择提供信息。与公共卫生相关的声明：我们的研究重点是口咽念珠菌病，这是一种艾滋病患者面临风险的感染。我们的方法将确定引起这种感染的病原体表达的高优先级治疗靶点。结果将会是 指导药物开发，将有助于将基础研究的重点放在与这种疾病直接相关的基因产品上。

DESCRIPTION (provided by applicant): Oropharyngeal candidiasis (OPC) is among the first clinical manifestations of AIDS. It occurs in 50-95% of HIV+ individuals during the progression to AIDS, and overall prevalence among AIDS patients is estimated by the CDC at 9-31%. This infection is caused primarily by Candida albicans, a commensal fungus that can cause mucosal and invasive infections. Our overall objective is to understand the C. albicans gene products that govern host interaction and contribute to pathogenesis of OPC. We will use a new gene expression assay technology, nanoString nCounter RNA measurements, to determine levels of C. albicans gene expression during infection in widely used a murine OPC model. Our preliminary results correlate well with prior patient sample and in vitro infection gene expression data, but also reveal extensive stress-responsive gene expression and rewiring of transcription factor-target gene relationships. Proposed studies focus on two classes of gene products: transcription factors, and surface and secreted proteins. We have chosen transcription factor genes because they are a highly regulated gene set, and their products have been or can be connected to functions through systematic mutant screens, expression profiling, and chromatin immunoprecipitation. We have chosen surface and secreted protein genes because their products are engaged in direct interaction with host cells, and because they present the most accessible therapeutic targets. We will develop a reference dataset that explores expression of these genes, and extend the analysis with functional validation. Our findings will generate new hypotheses and inform choices of genes and pathways that can be pursued by all investigators in the community. Statement of Public Health Relevance: Our studies focus on oropharyngeal candidiasis, an infection for which AIDS patients are at risk. Our approach will identify high-priority therapeutic targets expressed by the pathogen that causes this infection. The results will guide drug development and will help to focus basic research studies on gene products that are directly relevant to this disease.