Targeting the regulatory mechanism of hyphae to lateral yeast growth as a novel therapeutic approach against candidiasis

针对菌丝对侧向酵母生长的调节机制作为对抗念珠菌病的新治疗方法

• 批准号: 9817046
• 负责人: PRIYA UPPULURI
• 金额: $ 37.08万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-09 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9817046
• 关键词: Active Sites; Antifungal Agents; Attention; BRCA1 gene; BRCT Domain; Binding; Biological Assay; Blood Circulation; Candida; Candida albicans; Candidiasis; Carbon; Catheters; Cells; Coin; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Disease; Disseminated candidiasis; Distal; Down-Regulation; Drug resistance; Evaluation; FDA approved; Filament; Gene Deletion; Genes; Glucose; Goals; Growth; Human; Hyphae; In Vitro; Infection; Interruption; Invaded; Lateral; Lead; Libraries; Link; Mediating; Medical Device; Microbial Biofilms; Modeling; Morphogenesis; Mus; Mycoses; Organ; Pathogenesis; Pathway interactions; Patient-Focused Outcomes; Pharmaceutical Preparations; Pharmacology; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Play; Process; Production; Protein Biochemistry; Protein Dephosphorylation; Protein Inhibition; Proteins; Regulation; Role; Saccharomycetales; Septal Region; Signal Transduction; Specificity; Tissues; Virulence; Yeasts; analog; candidemia; high throughput screening; in vivo; inhibitor/antagonist; lead candidate; mouse model; novel; novel therapeutic intervention; novel therapeutics; pathogenic fungus; premature; prevent; protein protein interaction; ras Proteins; small molecule inhibitor; small molecule libraries; therapeutic target; trait

Abstract A single species Candida albicans, causes half of all invasive fungal infections in humans. The ability of C. albicans to switch reversibly between yeast and hyphe is a major virulence trait that helps it disseminate into the bloodstream (yeast) and invade target organs (filaments). Yeast to hyphae morphogenesis has been extensively studied and its regulation well understood. To the contrary, little is known about the reverse process: hyphae to yeast growth. C. albicans hyphae produce yeast cells from their lateral septal regions, coined as “lateral yeasts”. These lateral yeasts are always found with hyphae at the site of active infection, are the major cells that re- enter the bloodstream and establish distal foci of infection. In fact, lateral yeast cells released from the hyphal layers of biofilm-contaminated catheters have direct access into the bloodstream. We identified the first regulator of hyphae-to-lateral yeast growth, PES1 and have shown that blocking the process (by depleting PES1 in vivo) can abrogate disseminated candidiasis as well as biofilm- associated candidemia. Nothing is known on the regulatory aspects pf PES1. Our preliminary studies show that phosphorylation of Pes1 by Ras-PKA inhibits lateral yeast growth while its dephosphorylation by a Ras- linked phosphatase Yvh1 activates lateral yeast emergence from hyphae and induces biofilm dispersal. Using PES1 as a target for identifying small molecule inhibitors of lateral yeast growth, identified alexidine dihydrochloride that directly inhibited both Pes1 and Yvh1 and protected mice from biofilm-associated disseminated candidiasis. Here, using protein biochemistry assays, we propose to delineate how signaling through Ras-PKA regulates Pes1. We will identify other cognate regulators that interact with Pes1 to control lateral yeast growth, and use this information to discover novel compounds that can interrupt hyphae to lateral yeast growth and disseminated candidiasis. Ultimately, better outcomes for patients with indwelling medical devices is the goal of this application.

抽象的 单一物种白色念珠菌会引起人类所有侵入性真菌感染的一半。 C.的能力 白色唱片在酵母和菌丝之间可逆地切换是一种主要的病毒特征，可帮助其传播到 血液（酵母）和入侵目标器官（细丝）。酵母到菌丝形态发生已经 广泛的研究及其调节众所周知。与之形成鲜明对比的是，关于反面知之甚少 过程：菌丝到酵母生长。白色念珠菌菌丝从其侧隔区域产生酵母细胞， 创造为“侧酵母”。这些侧向酵母总是在活性感染部位发现有菌丝， 是重新进入血液并建立远端感染灶的主要细胞。实际上，外侧酵母 从生物膜污染导管的菌丝层释放的细胞已直接进入 血液。我们确定了第一个菌丝到外侧酵母生长的调节剂，PES1，并表明这是 阻止该过程（通过体内耗尽PES1）可以消除传播的念珠菌病以及生物膜 - 相关的候选血症。在调节方面尚无PF PES1。我们的初步研究表明 RAS-PKA对PES1的磷酸化抑制了横向酵母的生长，而其去磷酸化则通过Ras-磷酸化。 连接的磷酸酶YVH1激活了菌丝的外侧酵母出现，并诱导生物膜分散。使用 PES1作为鉴定侧向酵母生长的小分子抑制剂的靶标 直接抑制PES1和YVH1并保护与生物膜相关的小鼠的二氢氯化物 传播念珠菌病。在这里，使用蛋白质生物化学测定法，我们建议描述如何发出信号 通过RAS-PKA调节PES1。我们将确定其他与PES1相互作用以控制的同源调节器 横向酵母生长，并使用此信息发现可以中断菌丝到侧面的新型化合物 酵母生长和传播念珠菌病。最终，对于留置医疗的患者，更好的结果 设备是此应用程序的目标。