Regulation of Intrinsic Caspofungin Resistance in C. neoformans

新型隐球菌内在卡泊芬净耐药性的调节

基本信息

批准号：
9761969
负责人：
John C Panepinto
金额：
$ 22.56万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-08 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9761969
关键词：
5&apos Untranslated Regions Acquired Immunodeficiency Syndrome Amino Acid Sequence Amphotericin B Antifungal Agents Antifungal Therapy Arginine Binding C-terminal Caspofungin Cell Wall Cells Cellular Immunity Cessation of life Consensus Cryptococcosis Cryptococcus Cryptococcus neoformans Cryptococcus neoformans infection Data Defect Drug Targeting Elements Exhibits Fluconazole Flucytosine Foundations Fungal Proteins Future Genetic Genetic Transcription Hypersensitivity Immunocompromised Host In Vitro Industrial fungicide Lead Link Meningitis Messenger RNA Methylation Modeling Mutagenesis Mutation Pathogenicity Patients Pharmaceutical Preparations Polyadenylation Post-Transcriptional Regulation Predisposition Protein-Arginine N-Methyltransferase Proteins RNA Binding RNA Recognition Motif RNA-Binding Proteins Regulation Resistance Resources Ribosomes Role Site Testing Translating Translations Transplant Recipients Work echinocandin resistance glucan synthase in vivo inhibitor/antagonist mRNA Stability mortality mutant protein B resistance mechanism stressor

项目摘要

Abstract: Cryptococcosis causes hundreds of thousands of deaths per year, mostly in resource-poor settings. C. neoformans exhibits intrinsic resistance to echinocandin antifungals, eliminating this safe and effective class of antifungal drug from the anti-cryptococcal arsenal. The mechanism of resistance is unknown, but it is independent of drug target inhibition, as the target protein, the β-1,3-glucan synthase Fks1, is sensitive to the drug. We have identified a regulator of caspofungin sensitivity in C. neoformans, Puf4, that controls the FKS1 mRNA at the level of stability, and a puf4 mutant is caspofungin resistant. Deletion putative post-translational regulator of Puf4, the protein arginine methyltransferase (PRMT) Rmt5, confers hypersensitivity to caspofungin, suggesting a role for arginine methylation in the regulation of Puf4 function and caspofungin sensitivity. Our scientific premise is that Puf4 controls caspofungin sensitivity through post-transcriptional regulation of the FKS1 mRNA, and that the ability of Puf4 to regulate FKS1 is dependent on its methylation status. We will investigate the mechanism of Puf4 regulation including its binding to the FKS1 mRNA 5’ UTR and the effect of that interaction on translation and mRNA stability (Aim 1). We will then investigate the role of Puf4 methylation on its ability to regulate the FKS1 mRNA including the effect of rmt5 deletion on FKS1 post- transcriptional regulation, the identification of methylation sites on Puf4 and the use of mutagenesis to determine the consequence of methylation on effector function (Aim 2). PRMTs are known drug targets. Determining the mechanism by which Rmt5 and its putative target Puf4 control caspofungin sensitivity may lead to adjunctive therapies to potentiate the anti-cryptococcal activity of echinocandins.

抽象的：隐球菌病每年导致数十万人死亡，大部分发生在资源匮乏的地区。新型隐球菌表现出对棘白菌素抗真菌药物的内在耐药性，从而消除了这一类安全有效的真菌抗隐球菌库中的抗真菌药物的耐药机制尚不清楚，但确实如此。与药物靶标抑制无关，因为靶蛋白 β-1,3-葡聚糖合酶 Fks1 对我们已经鉴定出新型隐球菌中卡泊芬净敏感性的调节因子 Puf4，它控制着 FKS1。 mRNA 在稳定性水平上，puf4 突变体具有卡泊芬净推定的翻译后删除抗性。 Puf4 蛋白精氨酸甲基转移酶 (PRMT) Rmt5 的调节因子，赋予对卡泊芬净，表明精氨酸甲基化在调节 Puf4 功能和卡泊芬净中的作用我们的科学前提是 Puf4 通过转录后控制卡泊芬净敏感性。 Puf4 调节 FKS1 mRNA 的能力取决于其甲基化我们将研究 Puf4 调节机制，包括其与 FKS1 mRNA 5' UTR 的结合。以及这种相互作用对翻译和 mRNA 稳定性的影响（目标 1）。 Puf4 甲基化对其调节 FKS1 mRNA 的能力的影响，包括 rmt5 缺失对 FKS1 后的影响转录调控、Puf4 上甲基化位点的鉴定以及使用诱变确定甲基化对效应器功能的影响（目标 2）。确定 Rmt5 及其假定靶点 Puf4 控制卡泊芬净敏感性的机制导致辅助疗法增强棘白菌素的抗隐球菌活性。