HOST FACTORS IN CRYPTOCOCCAL INFECTION

描述（由申请方提供）：新型隐球菌是一种机会致病菌，可导致艾滋病或其他免疫功能低下患者发生危及生命的疾病。该R21应用程序的重点是这种兼性细胞内病原体与其哺乳动物宿主的吞噬细胞的相互作用。这些相互作用会对宿主和酵母造成不同程度的损害，但所涉及的机制知之甚少，限制了我们影响有利于宿主的解决方案的能力。我们建议将联合收割机RNA干扰技术和高通量筛选技术相结合，以鉴定C.新生物进入宿主细胞目的一是优化评价C.宿主细胞受到RNA干扰的新生儿。目的II是通过筛选靶向已知基因的RNAi文库来鉴定真菌粘附和进入所需的序列，并确认和优先考虑这些序列用于进一步的研究。结果将使用严格的统计方法和适当的控制进行评估，并将在我们在隐球菌及其宿主生物学方面的专业知识的背景下进行解释。我们研究的长期目标是了解隐球菌病的发病机制。拟议的研究将阐明决定生物体存活和传播以及宿主限制疾病能力的发病机制的一个关键方面。这项工作还将为重要病原体的研究开辟新的领域，并开发可随时应用于病原体生物学其他领域的实验工具。公共卫生相关性：这项研究与公共卫生高度相关，因为所研究的生物体会导致严重的人类疾病，而目前的治疗方法还不足以治疗这些疾病。新型隐球菌与宿主细胞的相互作用是其致病能力的基础，因此了解和抑制这些过程可能会促进隐球菌感染的治疗。此外，这项工作将有助于基础科学知识，这将提供对病原微生物以及其他生物学领域的见解。

DESCRIPTION (provided by applicant): Cryptococcus neoformans is an opportunistic pathogen that is responsible for life-threatening disease in patients with AIDS or other conditions of immunocompromise. This R21 application focuses on the interactions of this facultative intracellular pathogen with the phagocytic cells of its mammalian host. These interactions resolve with varying levels of damage to the host and yeast, but the mechanisms involved are poorly understood, limiting our ability to influence resolution in favor of the host. We propose to combine RNA interference methodology and high-throughput screening assays to identify host factors that are required for the entry of C. neoformans into host cells. Aim I is to optimize methods for assessing the interactions of C. neoformans with host cells subjected to RNA interference. Aim II is to identify sequences required for fungal adherence and entry by screening RNAi libraries targeting known genes, and to confirm and prioritize these sequences for additional investigation. Results will be assessed using rigorous statistical methods and appropriate controls, and will be interpreted in the context of our expertise in the biology of Cryptococcus and its host. The long-term goal of our studies is to understand the pathogenesis of cryptococcosis. The proposed studies will elucidate a crucial aspect of pathogenesis that determines survival and dissemination of the organism and the host's ability to limit disease. This work will additionally open new areas for investigation of an important pathogen and develop experimental tools that can be readily applied to other areas of pathogen biology. PUBLIC HEALTH RELEVANCE: This research is highly relevant to public health because the organism under study causes serious human illness for which current therapies are not adequate. The interactions of Cryptococcus neoformans with host cells are fundamental to its ability to cause disease, so understanding and inhibiting these processes may advance treatment of cryptococcal infection. Further, this work will contribute to basic science knowledge, which will provide insights into pathogenic microbes as well as other areas of biology.