Fungal and Host Factors in Initiation of Cryptococcal Persistence

项目摘要 候选人我的职业目标是改善传播性传染病的检测和管理选择， 通过研究宿主与微生物的相互作用来预防疾病。我打算通过终身教职来实现这一目标 作为初级教师，有大量的保护时间进行基础研究。在短期内，我的目标是扩大我的 作为一名调查员的技能，并为我过渡到独立研究奠定基础。教育 该提案的各个方面围绕实验真菌学和基因表达分析， 将使我能够提高我们对来自宿主和病原体的播散性真菌感染的理解 视角 Research.隐球菌脑膜脑炎是一种毁灭性的疾病， 免疫功能低下的患者，如艾滋病或移植患者。脑部感染很难治疗， 发生在初次手术后数月甚至数年，通常是不被注意的肺部感染。因此，真菌 被认为在健康个体中长期处于静止状态，而在这种状态下， 似乎对抗真菌治疗敏感更好地理解静止状态，它是如何启动的 和维护，应该为我们提供更好的检测和管理工具，并给我们机会， 在它引发致命疾病之前控制这种静止的感染。 斑马鱼幼虫作为模式宿主，可以直接监测体内微生物的发病机制。相比之下 在细胞培养模型中的发现，在斑马鱼中，我们发现接种的隐球菌可以有多种命运， 包括清除、复制和散布或吞噬细胞内的静止。这种静止状态， 只要幼虫存活，就能持续很长时间，孢子感染后很常见，但当幼虫 接种酵母菌。我将使用孢子感染作为早期静止感染的模型。拟议 实验的目的是定义允许持久性的主机响应的功能，并定义 隐球菌细胞诱导这种反应的状态。 职业发展计划。在拟议的研究期间，我的主要目标将是发扬我的 以前在隐球菌感染的先天免疫方面的工作，同时将真菌生物学的考虑加入到我的研究中。 接近疾病。我还将获得基因表达分析，生物统计学和真菌 生物学通过指导，动手实验台研究和教学式学习。沿着，我计划开发新的 领导、指导和管理技能，使我能够向独立过渡。我计划协调 我的研究和培训活动，以建立一个成功的R01应用程序在第四年的奖项。 环境我将在威斯康星大学麦迪逊分校的儿科得到很好的支持。 该机构充满了不同背景和兴趣的科学和医学教师， 计算、生物技术、实验动物护理和显微镜的基础设施都很好。

PROJECT SUMMARY Candidate. My career goal is to improve the detection and management options for disseminated infectious diseases through investigation of host-microbe interactions. I plan to pursue this with a tenure-track appointment as junior faculty with significant protected time for basic research. In the short term, my goal is to broaden my skills as an investigator and build the foundation for my transition to independent research. The education aspects of this proposal center around experimental mycology and gene expression analysis, new skills which will enable me to improve our understanding of disseminated fungal infections from both host and pathogen perspectives. Research. Cryptococcal meningoencephalitis is a devastating disease most common in the immunocompromised, such as AIDS or transplant patients. The brain infection is very difficult to treat and can occur months or even years after the initial exposure—often an unnoticed pulmonary infection. Thus, the fungus is thought to exist in a quiescent state for long periods in healthy individuals, and while in this state does not appear to be susceptible to antifungal treatment. A better understanding of the quiescent state, how it is initiated and maintained, should provide us with better detection and management tools and give us the chance to manage this quiescent infection before it causes a deadly disease. The zebrafish larva as a model host allows direct monitoring of microbial pathogenesis in vivo. In contrast to findings in cell culture models, in the zebrafish we find that inoculated Cryptococcus can have multiple fates, including clearance, replication and dissemination, or quiescence within phagocytes. This quiescent state, which can last for as long as long as the larva is viable, is common after spore infection but rare when larvae are inoculated with the yeast form. I will use spore infection as a model for early quiescent infection. The proposed experiments are designed to define the features of the host response that allow for persistence, and to define the state of cryptococcal cells that induce this response. Career Development Plan. During the proposed research period, my primary goal will be to carry forward my previous work in innate immunity to cryptococcal infection while adding considerations of fungal biology to my approach to the disease. I will also obtain new training in gene expression analysis, biostatistics and fungal biology through mentoring, hands-on bench research and didactic learning. Along with this I plan to develop new skills in leadership, mentoring and management to enable my transition to independence. I plan to coordinate my research and training activities to build toward a successful R01 application in the fourth year of the award. Environment. I will be well supported in the Department of Pediatrics at the University of Wisconsin-Madison. The institution is replete with a scientific and medical faculty of diverse backgrounds and interests, and the infrastructure for computing, biotechnology, laboratory animal care and microscopy are excellent.