Fungal and Host Factors in Initiation of Cryptococcal Persistence

项目总结 候选人。我的职业目标是改进对传播性传染病的检测和管理 通过研究宿主-微生物的相互作用来防治疾病。我计划通过终身教职任命来实现这一点。 作为初级教员，他们有大量受保护的基础研究时间。在短期内，我的目标是扩大我的 作为一名研究人员的技能，并为我过渡到独立研究奠定了基础。教育 这项提案的各个方面都围绕着实验真菌学和基因表达分析，这些新技术 将使我能够更好地了解来自宿主和病原体的播散性真菌感染 透视。 研究。隐球菌性脑膜脑炎是一种破坏性疾病，最常见于 免疫功能受损，如艾滋病或移植患者。脑部感染很难治疗，而且可以 在初次接触几个月甚至几年后发生--通常是一种未被注意到的肺部感染。因此，这种真菌 被认为在健康个体中长时间处于静止状态，而在这种状态下则不存在 似乎对抗真菌治疗敏感。更好地理解静止状态，以及它是如何启动的 并进行维护，应该为我们提供更好的检测和管理工具，并使我们有机会 在它导致致命疾病之前，控制这种静止的感染。 斑马鱼幼体作为模型宿主，可以直接监测体内的微生物致病情况。相比之下， 根据在细胞培养模型中的发现，在斑马鱼中，我们发现接种了隐球菌可以有多种命运， 包括清除、复制和传播，或吞噬细胞内的静止。这种静止状态， 只要幼虫还活着，就能存活多久，这在孢子感染后很常见，但当幼虫 接种酵母菌。我将用孢子感染作为早期静止性感染的模型。建议数 实验旨在定义允许持久化的主机响应的特征，并定义 引起这种反应的隐球菌细胞的状态。 职业发展计划。在拟议的研究期间，我的主要目标将是推进我的 对隐球菌感染的天然免疫方面的先前工作，同时增加了对真菌生物学的考虑 这种疾病的治疗方法。我还将接受基因表达分析、生物统计学和真菌方面的新培训 通过指导、实际操作的板凳研究和授课学习来学习生物学。随之而来的是我计划开发新的 领导、指导和管理方面的技能使我能够过渡到独立。我计划协调 我的研究和培训活动为R01的成功申请奠定了基础，在该奖项的第四年。 环境我会在威斯康星大学麦迪逊分校的儿科得到很好的支持。 该机构拥有一批具有不同背景和兴趣的科学和医学教员， 计算机、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.