Investigation on Replicative aging in Cryptococcus neoformans populations

新型隐球菌种群复制衰老的研究

• 批准号: 10867739
• 负责人: Bettina Fries
• 金额: $ 55.51万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-27 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10867739
• 关键词: ATP-Binding Cassette Transporters; Abbreviations; Acidity; Acquired Immunodeficiency Syndrome; Affect; Age; Aging; Amphotericin B; Anabolism; Antifungal Agents; Antifungal Therapy; Brain; Calcium; Cell Wall; Cells; Cerebrospinal Fluid; Chronic; Clinical; Cloning; Cryptococcus; Cryptococcus neoformans; Cryptococcus neoformans infection; Data; Devices; Electrons; Ergosterol; Fluconazole; Funding; Generations; Glucose; Heterogeneity; Human; Hydrogen Peroxide; Immunocompromised Host; Infection; Interferometry; Intermediate resistance; Investigation; Knowledge; Laboratories; Link; Longevity; Macrophage; Matrix Metalloproteinases; Membrane Potentials; Meningoencephalitis; Microdissection; Microfluidic Microchips; Microscopic; Mitochondria; Morbidity - disease rate; Morphology; Mothers; Mycoses; Names; Pathogenesis; Patients; Persons; Phagocytes; Phagolysosome; Pharmaceutical Preparations; Phenotype; Physiological; Play; Population; Process; Publishing; Reactive Oxygen Species; Research; Research Proposals; Resistance; Role; Saccharomyces cerevisiae; Sampling; Starvation; Stress; System; Techniques; Transmission Electron Microscopy; Treatment Failure; Up-Regulation; Vacuole; Yeasts; chronic infection; drug development; efflux pump; extracellular vesicles; high throughput screening; in vivo; mathematical model; metabolomics; microscopic imaging; mitochondrial dysfunction; mitochondrial membrane; mortality; mutant; novel; overexpression; pathogen; pathogenic fungus; programs; protein expression; rapid diagnosis; resilience; synergism; tool; trafficking; vesicle transport; virulence gene

This application is a competitive renewal of a research proposal that investigated replicative aging in Cryptococcus neoformans, a human pathogenic fungus that is a major cause of mortality and morbidity in AIDS patients. Replicative aging leads to old C. neoformans cells, which are more resistant to clearing by the host cells, as well as antifungal treatment. The proposed studies are focused on continuing our prior studies by establishing mechanisms that convey the resilience of these old C. neoformans cells. The overarching hypothesis is that the observed resistance to phagocytic attack and antifungal drugs results from a combination of several physiological changes that occur in C. neoformans cells during replicative aging. These changes occur early in the replicative lifespan, and they synergize to further the selection and accumulation of old C. neoformans cells. In the past funding period, we made significant progress, including developing a high- throughput microfluidic chip device, which now enabled us to study morphology and protein expression in C. neoformans populations. Other discoveries obtained in the last funding cycle include knowledge about cell wall remodeling, altered mitochondrial function, and cloning of a novel efflux pump that is expressed in old C. neoformans cells. These findings now guide the proposed experimental approach. Four aims, each supported by comprehensive preliminary and published data, are proposed. Aim 1 will establish mechanisms how old C. neoformans cells are selected by macrophages. Aim 2 will seek to establish how altered mitochondrial function in old C. neoformans cells affects tolerance to antifungal drugs. In Aim 3, we propose to clone a mother enrichment mutant. Aim 4 will establish how vacuolar function of old C. neoformans cells affects tolerance to antifungal drugs.

这项申请是对一项研究方案的竞争性更新，该研究方案研究了 新生隐球菌，一种人类致病真菌，是艾滋病死亡和发病的主要原因 病人。复制性衰老导致新生葡萄球菌细胞老化，它们对宿主的清除更具抵抗力 细胞，以及抗真菌治疗。拟议研究的重点是继续我们先前的研究，包括 建立机制来传递这些旧的新生隐孢子虫细胞的弹性。最重要的是 假说是，观察到的对吞噬攻击和抗真菌药物的抵抗力是联合作用的结果。 在复制衰老过程中，新生隐孢子虫细胞中发生的几种生理变化。这些变化 发生在复制寿命的早期，它们协同作用进一步选择和积累旧的C. 新生细胞。在过去的资助期间，我们取得了重大进展，包括开发了一种 吞吐量微流控芯片设备，这使得我们能够研究C。 新形态菌种群。在上一个资助周期中获得的其他发现包括关于细胞壁的知识 重塑，线粒体功能改变，以及一个新的外排泵的克隆，这是在旧的C. 新生细胞。这些发现现在指导了拟议的实验方法。四个目标，每个目标都得到支持 通过综合的初步数据和已发表的数据，提出了一些建议。目标1将建立C。 新生细胞是由巨噬细胞选择的。目标2将寻求确定改变的线粒体功能如何 在旧的新生葡萄球菌中，细胞会影响对抗真菌药物的耐受性。在目标3中，我们提议克隆一位母亲 浓缩突变体。目标4将确定新生隐孢子虫旧细胞的空泡功能如何影响对 抗真菌药物。