GENETIC IDENTIFICATION OF ANTIFUNGAL DRUG TARGETS

抗真菌药物靶点的基因鉴定

• 批准号: 6217114
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6217114
• 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.

该提案有三个具体的短期目标：（1）。克隆C. 可能对发病机制重要的新生儿基因; 2.） 在这些基因中构建“敲除”突变; nd 3.）测试 结果C.实验性感染中的新型突变体的目标 这些实验是为了鉴定无毒/无毒的基因突变体， 实验感染中的致病性，从而确定良好的 潜在的药物靶点 主要关注的是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， 许多营养缺陷型是无毒的。这些途径中基因的选择是 （1）由于（1）它们在人类中的缺失和（2）高度有害的 特定突变体的体外表型。第二个重点是基因 只有在高温下才能生长的产品。的 关注这些基因产物的原因是， 在高温下是真菌致病力所必需的。 该提案与健康有关，因为（i）它涉及C。新生动物， 最严重的致病真菌之一，（ii）它是集中在 抗真菌药物靶点的鉴定。当前 可用的抗真菌药物很少（与抗菌药物相比）， 数量，比临床医生想要的效果要差，并且往往 严重的副作用。因此，非常需要开发新的 抗真菌药这项提案的目标是从基因上识别新的 抗真菌药物靶点。一旦确定了具体的新目标， 将有可能把注意力集中在这些基因产物和用途上， 新的技术，如组合化学，以开发小 分子抑制剂。这样的合作在杜克是可能的（见 附信）。 该提案有三个广泛的长期/未来目标：1。审查 更多的基因/突变体，以获得C.新生 发病机理; 2.）将这种方法应用于其他病原真菌;以及 3.）第三章开发基因产物的小分子抑制剂， 对发病机理至关重要。