Molecular Mechanisms that Regulate Phase Transition in Dimorphic Fungi

调节二态真菌相变的分子机制

基本信息

批准号：
7921227
负责人：
Gregory M Gauthier
金额：
$ 5万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-17 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7921227
关键词：
Agrobacterium Antifungal Agents Biological Models Blastomyces dermatitidis Breathing Candidate Disease Gene Cell Wall Chitin Complement Complementary DNA Cyclic AMP Cysteine Cytokinesis DNA Defect Disease Drug Delivery Systems Environmental Risk Factor Epidemic Foundations Gene Expression Gene Family Gene Transfer Genes Genetic Genome Glucans Goals Hand Host Defense Host Defense Mechanism Immunity Infection Insertional Mutagenesis Lead Lung Master&apos s Degree Mediating Medical Mentors Modeling Molds Molecular Molecular Biology Molecular Genetics Monitor Morphogenesis Morphology Mus Mutation Mycelium Nitrogen Nutrient Orthologous Gene Pathogenesis Phase Phase Transition Phenotype Predisposition Public Health RNA Interference Regulation Research Research Personnel Research Training Rhizobium radiobacter Role Scientist Signal Transduction Solid Starvation Temperature Testing The science of Mycology Training Programs Virulence Work Yeasts clinical practice fungus gene discovery gene function in vivo insight interest knock-down member microbial mortality mutant novel pathogen phosphoric diester hydrolase positional cloning programs skills tool

项目摘要

DESCRIPTION (provided by applicant): The endemic dimorphic fungi are increasingly significant pathogens in clinical practice and cause substantial mortality in those with impaired immunity. These agents must convert from mold to yeast to acquire virulence and escape host defenses. Despite the pivotal role of morphogenesis in pathogenesis of these infections, the mechanisms that regulate the phase transition are poorly understood. The goal of my research is to uncover and characterize genes that regulate phase transition in dimorphic fungi using Blastomyces dermatitidis as a model. I hypothesize that genes involved in environmental sensing, intracellular signaling, cytokinesis, and morphology govern phase transition. To investigate this, I propose two aims with complementary approaches, using forward and reverse genetic strategies: 1) Elucidate the identity and function of genes in B. dermatitidis mutants defective in phase transition that I have already generated by Agrobacterium-mediated DNA transfer, and 2) Investigate the role in morphogenesis of two orthologs in the B. dermatitidis genome postulated to be involved in fungal phase transition - septin and phosphodiesterase - by knocking down their expression with RNA interference. My experimental approach will involve the use of cutting-edge molecular biology tools to discover new genes, discern their role in morphogenesis, and test their impact on virulence in a murine model of pulmonary infection. The results of my research will provide fundamental new insight about the molecular mechanisms behind phase transition in dimorphic fungi. The research training proposed will allow me to hone the skills I will need as a bench scientist to advance the field of molecular medical mycology. The coursework will give me a solid intellectual foundation in molecular genetics, microbial pathogenesis and medical mycology, while fulfilling the requirements for a Masters Degree. A mentoring committee with expertise in fungal pathogenesis will guide my progress in the lab and classroom. The overall training program will allow me to establish a contemporary research program investigating molecular regulation of morphogenesis in dimorphic fungi. The research proposed is relevant to public health because it will provide insight into the mechanisms by which fungi cause disease and may lead to the identification of new antifungal drug targets.

描述（由申请人提供）：流行的二态真菌在临床实践中越来越重要，在免疫力受损的人中导致大量死亡。这些代理必须从霉菌转换为酵母，以获得毒力并逃脱宿主防御力。尽管形态发生在这些感染的发病机理中的关键作用，但调节相变的机制却很少了解。我的研究的目的是使用Blastomyces Dermatitidis作为模型来揭示和表征调节双态真菌中相变的基因。我假设与环境传感，细胞内信号，细胞因子和形态学有关的基因控制相变。 To investigate this, I propose two aims with complementary approaches, using forward and reverse genetic strategies: 1) Elucidate the identity and function of genes in B. dermatitidis mutants defective in phase transition that I have already generated by Agrobacterium-mediated DNA transfer, and 2) Investigate the role in morphogenesis of two orthologs in the B. dermatitidis genome postulated to be involved in fungal phase transition - septin和磷酸二酯酶 - 通过用RNA干扰击倒其表达。我的实验方法将涉及使用尖端的分子生物学工具来发现新基因，辨别它们在形态发生中的作用，并在肺部感染的鼠模型中测试其对毒力的影响。我的研究结果将为二态真菌中相变背后的分子机制提供基本的新见解。提出的研究培训将使我能够磨练我作为替补科学家所需的技能，以推动分子医学真菌学领域。该课程将使我在分子遗传学，微生物发病机理和医学真菌学方面为我提供了坚实的知识基础，同时满足了硕士学位的要求。具有真菌发病机理专业知识的指导委员会将指导我在实验室和教室中的进步。整个培训计划将使我能够建立一个当代研究计划，研究二态真菌中形态发生的分子调节。提出的研究与公共卫生有关，因为它将洞悉真菌引起疾病的机制，并可能导致鉴定新的抗真菌药物靶标。