GENETIC IDENTIFICATION OF ANTIFUNGAL DRUG TARGETS

抗真菌药物靶点的基因鉴定

• 批准号: 6299763
• 负责人: John H. McCusker
• 金额: $ 15.15万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6299763
• 关键词: Cryptococcus neoformans; aminoacid biosynthesis; antifungal agents; cryptococcosis; disease /disorder model; fungal genetics; gene expression; gene mutation; gene targeting; laboratory mouse; microorganism growth; molecular cloning; virulence; vitamin biosynthesis

This proposal has three specific, short term aims: 1.) cloning C. neoformans genes likely to be important for pathogenesis; 2.) constructing "knock-out" mutations in these genes; nd 3.) testing the resulting C. neoformans mutants in experimental infections. The goal of these experiments is to identify genes-mutants that are avirulent/non- pathogenic in experimental infections and thereby identify good potential drug targets. The primary focus is on a specific subset of amino acid and vitamin biosynthetic genes. The reason for focusing on these pathways is that many auxotrophs are avirulent. The choice of genes in these pathways is dictated by (i) their absence in humans and (ii) the highly deleterious in vitro phenotypes of specific mutants. The secondary focus is on gene products that are required for growth only at high temperature. The reason for focusing on such gene products is that the ability to growth at high temperatures is essential for fungal virulence-pathogenesis. This proposal is health related because (i) it deals with C. neoformans, one of the most serious of the pathogenic fungi, and (ii) it is focused on the identification of antifungal drug targets. The currently available antifungal drugs are few (compared to antibacterials) in number, are less effective than clinicians would like, and tend to have severe side effects. Therefore, there is a great need to develop new antifungals. The goal of this proposal is to genetically identify new antifungal drug targets. Once specific new targets have been identified it will be possible to focus attention on these gene products and use novel technologies, such as combinatorial chemistry, to develop small molecule inhibitors. Such collaborations are possible here at Duke (see attached letters). This proposal has three broad long term/future goals: 1.) to examine more genes/mutants to get a global picture of C. neoformans pathogenesis; 2.) to apply this approach to other pathogenic fungi; and 3.) to develop small molecule inhibitors of gene products that are essential for pathogenesis.

该提案具有三个具体的短期目标：1.) 克隆 C. 新型隐球菌基因可能对发病机制很重要； 2.) 在这些基因中构建“敲除”突变； 3.) 测试 实验感染中产生的新型隐球菌突变体。目标是 这些实验是为了鉴定无毒/无毒的基因突变体 实验感染中具有致病性，从而鉴定出良好的 潜在的药物靶点。 主要关注氨基酸和维生素的特定子集 生物合成基因。关注这些途径的原因是 许多营养缺陷型细菌是无毒的。这些途径中基因的选择是 取决于（i）它们在人类中不存在以及（ii）高度有害 特定突变体的体外表型。第二个重点是基因 仅需要在高温下生长的产品。这 关注此类基因产品的原因是生长能力 高温下对于真菌毒力发病机制至关重要。 该提案与健康相关，因为 (i) 它涉及新型隐球菌， 最严重的病原真菌之一，并且 (ii) 它是重点关注的 抗真菌药物靶点的鉴定。目前的 可用的抗真菌药物很少（与抗菌药物相比） 数量，效果不如临床医生希望的那样，并且往往有 severe side effects.因此，非常需要开发新的 抗真菌剂。该提案的目标是从基因上鉴定新的 抗真菌药物靶点。一旦确定了具体的新目标 将有可能将注意力集中在这些基因产物和使用上 新技术，如组合化学，开发小型 分子抑制剂。这种合作在杜克大学是可能的（参见 附信）。 该提案具有三个广泛的长期/未来目标：1.) 审查 更多基因/突变体以获得新型隐球菌的全球概况 发病; 2.)将此方法应用于其他病原真菌；和 3.) 开发基因产物的小分子抑制剂 发病机制所必需的。