CRYPTOCOCCAL INTERACTIONS WITH HOST CELLS

隐球菌与宿主细胞的相互作用

• 批准号: 8038303
• 负责人: Tamara L Doering
• 金额: $ 37.24万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8038303
• 关键词: Acquired Immunodeficiency Syndrome; Adherence; Affect; Agrobacterium; Area; Basic Science; Biochemical; Biological Assay; Biology; Cell Adhesion; Cells; Cryptococcus; Cryptococcus neoformans; Cryptococcus neoformans infection; Disease; Drug resistance; Event; Exhibits; Extracellular Fluid; Gene Deletion; Genes; Genetic; Goals; Growth; Health; Human; Immunocompromised Host; Individual; Infection; Insertional Mutagenesis; Investigation; Knowledge; Libraries; Life; Mediating; Meningoencephalitis; Methods; Microbe; Molecular; Organism; Pathogenesis; Patients; Phagocytes; Population; Process; Proteins; Public Health; Recovery; Research; Resolution; Role; Screening procedure; Sequence Analysis; Severity of illness; Statistical Methods; System; Techniques; Virulence; Work; Yeasts; base; experience; fascinate; follow-up; fungus; high throughput screening; insight; mutant; pathogen; research study; tool

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; extrapulmonary cryptococcal infection is an AIDS-defining illness. The long-term goal of our studies is to understand the pathogenesis of cryptococcosis. This application focuses on a fundamental part of this process, 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 forward genetics, high-throughput screening assays, and focused follow-up studies to determine the cryptococcal factors that are required for this pathogen to successfully adhere to and enter host cells. Aim I is to identify mutants in these processes, using insertional mutagenesis and automated screening. Aim II is to determine the mechanisms by which selected gene products influence pathogen adherence and entry interactions. These studies will employ assays of cell adhesion, entry, and intracellular survival and growth; molecular techniques for sequence recovery, analysis, and gene deletion; and phenotypic assessment including morphological studies, biochemical analysis, and virulence assays. Results will be assessed using rigorous statistical methods and appropriate controls, and will be interpreted in the context of our expertise in the fundamental biology of Cryptococcus. The proposed studies will elucidate a crucial aspect of pathogenesis that determines the 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 experiment tools that can be applied to other areas of fungal 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.

描述（由申请人提供）：新甲壳虫是一种机会性病原体，负责艾滋病患者或其他免疫功能疾病患者的威胁生命疾病；肺外隐球菌感染是一种定义艾滋病的疾病。我们研究的长期目标是了解隐球菌病的发病机理。该应用的重点是该过程的基本部分，该过程的相互作用与哺乳动物宿主的吞噬细胞的相互作用。这些相互作用与对宿主和酵母的损害程度不同，但是所涉及的机制却鲜为人知，从而限制了我们影响分辨率而倾向于宿主的能力。我们建议将正向遗传学，高通量筛选分析和集中的后续研究结合起来，以确定该病原体成功遵守并进入宿主细胞所需的加密摄氏因子。目的I是使用插入诱变和自动筛选在这些过程中识别突变体。 AIM II是确定所选基因产物影响病原体依从性和进入相互作用的机制。这些研究将采用细胞粘附，进入以及细胞内生存和生长的测定。用于序列恢复，分析和基因缺失的分子技术；和表型评估，包括形态学研究，生化分析和毒力测定。结果将使用严格的统计方法和适当的控制来评估，并将在我们在加密赛基本生物学方面的专业知识的背景下进行解释。拟议的研究将阐明发病机理的关键方面，该方面决定了生物体的生存和传播以及宿主限制疾病的能力。这项工作还将为研究重要病原体的新领域开放，并开发可应用于其他真菌生物学领域的实验工具。公共卫生相关性：这项研究与公共卫生高度相关，因为研究中的生物会导致严重的人类疾病，因为当前疗法不足以适合。加密赛车与宿主细胞的相互作用是引起疾病能力的基础，因此理解和抑制这些过程可能会推动对隐球菌感染的治疗。此外，这项工作将有助于基础科学知识，这将提供对致病微生物以及其他生物学领域的见解。