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：加密环球每年会导致数十万人死亡，主要是在资源贫乏的环境中。 C Neoformans表现出对Echinocandin抗真菌剂的内在抗性，消除了这种安全有效的类别来自抗晶局武库的抗真菌药物。抵抗的机制尚不清楚，但这是独立于药物靶标，作为靶蛋白，β-1,3-葡聚糖合酶FKS1，对药品。我们已经确定了C. neoformans puf4中Caspofungin灵敏度的调节剂，该调节器控制FKS1 稳定水平的mRNA和PUF4突变体具有抗Caspofungin。删除推定后翻译 PUF4的调节剂，蛋白精氨酸甲基转移酶（PRMT）RMT5，赋予对超敏反应的 caspofungin，提示精氨酸甲基化在PUF4功能和caspofungin调节中的作用灵敏度。我们的科学前提是，PUF4通过转录后控制Caspofungin敏感性 FKS1 mRNA的调节，并且PUF4调节FKS1的能力取决于其甲基化地位。我们将研究PUF4调节的机制，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（AIM 1）。然后，我们将调查 PUF4甲基化对其调节FKS1 mRNA的能力，包括RMT5缺失对FKS1的影响转录调节，鉴定PUF4上的甲基化位点以及使用诱变确定甲基化对效应函数的后果（AIM 2）。 PRMT是已知的药物靶标。确定RMT5及其推定的目标PUF4控制Caspofungin灵敏度的机制可能导致辅助疗法可潜在的棘突蛋白的抗晶状体活性。