Pathobiology of C. neoformans in the Central Nervous System

中枢神经系统新型隐球菌的病理学

• 批准号: 7559532
• 负责人: John R. Perfect
• 金额: $ 39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7559532
• 关键词: Animal Model; Antifungal Agents; Antifungal Therapy; Attenuated; British Columbia; Central Nervous System Infections; Cerebrospinal Fluid; Cryptococcus; Cryptococcus neoformans; Cues; Data; Disease; Disease Marker; Disease Outbreaks; Disease Outcome; Encapsulated; Environment; Enzymes; Gene Expression; Gene Silencing; Gene Targeting; Genes; Genetic; Genomics; Growth; High temperature of physical object; Human; Immune; Immunity; Immunocompromised Host; In Vitro; Individual; Infection; Investigation; Island; Knowledge; Laboratories; Lead; Life; Link; Microarray Analysis; Mitochondria; Molecular; Morbidity - disease rate; Mutagenesis; Neuraxis; Oryctolagus cuniculus; Oxidative Stress; Pathogenesis; Pathway interactions; Pattern; Pharmaceutical Preparations; Phenotype; Population; Production; Proteins; Protocols documentation; Regulation; Respiration; Reverse Transcriptase Polymerase Chain Reaction; Role; Screening procedure; Signaling Pathway Gene; Site; Spottings; Stress; Subtraction Technique; Techniques; Technology; Temperature; Testing; Vaccines; Validation; Virulence; Virulent; Yeasts; base; cDNA Subtraction; congenic; design; functional genomics; fungus; genome sequencing; improved; in vivo; insight; mRNA Differential Displays; mortality; mutant; novel therapeutics; pathogen; reconstitution; response; serial analysis of gene expression

DESCRIPTION (provided by applicant): Cryptococcus neoformans is one of the most common life-threatening central nervous system infections in humans and despite present treatments, morbidity and mortality still remain high. In a recent outbreak on Vancouver Island, British Columbia it was shown that cryptococcal strains have evolved rapidly to produce more virulent strains. The focus of this proposal is to use molecular techniques to identify, characterize, and validate genes in C. neoformans which are important to the virulence composite of this encapsulated yeast. Our primary hypothesis is based around the simple assumption that under certain environmental stresses, C. neoformans strains will regulate their genetic networks/pathways for production of proteins to allow them to survive and grow in the hostile environment of the host. This proposal details a plan of investigations which allows use of new technological advances to perform a highly integrative but global screen of transcriptional profiling in order to predict the weak points in Cryptococcus pathobiology. Our laboratory has over the last few years validated that this overall strategy can be successfully used. We have used cDNA subtraction techniques, differential display RT-PCR, in vivo expression technology, serial analysis of gene expression (SAGE), and most recently microarray technology to successfully begin to identify genes and which make C. neoformans, a pathogen. Through our studies we have identified the following genes and pathways to be important to the virulence composite: 1) genes encoding enzymes; 2) oxidative stress genes; 3) signaling pathway genes; 4) high temperature growth genes; 5) impact of mitochondria and respiration; 6) influence of attenuated null mutants on host immunity. A cornerstone of this proposal is to use microarrays to harness their ability to collect massive data and yet attempt to focus our understanding to relevant genes and networks by using specific in vitro and vivo conditions. This screening design will be carefully linked to functional studies through creation of null mutants for phenotypic analysis in relevant animal models. Our overall scientific plan is to understand the genetic regulation of the virulence composite which will contain a powerful insight into discovery of gene targets that interrupt pathogenesis and lead to new therapeutic strategies such as antifungal drugs and vaccines. PROJECT NARRATIVE This project uses genomic strategies to capture what makes the life-threatening fungus Cryptococcus neoformans such an effective pathogen in immunocompromised individuals. With the knowledge gained from these studies it is anticipated that new antifungal targets can be identified to help develop new drugs and/or vaccines against this common fungal pathogen for an enlarging severely immunocompromised population.

描述（由申请人提供）：新型隐球菌是人类最常见的危及生命的中枢神经系统感染之一，尽管目前的治疗方法，发病率和死亡率仍然很高。最近在不列颠哥伦比亚省温哥华岛爆发的一次疫情表明，隐球菌菌株迅速进化，产生了更毒的菌株。本研究的重点是利用分子技术来鉴定、表征和验证新生酵母中对这种被封装酵母的毒力复合物很重要的基因。