Fungal calcium channels as therapeutic targets for AIDS-associated opportunistic

真菌钙通道作为艾滋病相关机会性感染的治疗靶点

• 批准号: 8015377
• 负责人: ANGIE GELLI
• 金额: $ 37.17万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8015377
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adjuvant; Anabolism; Antifungal Agents; Attenuated; Biological; Biological Assay; Calcium Channel; Candida albicans; Cell Line; Cells; Complex; Coupled; Cryptococcal Meningitis; Cryptococcus neoformans; Cryptococcus neoformans infection; Development; Doctor of Philosophy; Drug Combinations; Drug Compounding; Economics; Endoplasmic Reticulum; Environment; Equilibrium; Ergosterol; Event; Fluorescence Resonance Energy Transfer; Fungal Meningitis; Growth; Health; Image; Immune response; Immunosuppression; In Vitro; Industrial fungicide; Lead; Life; Maintenance Therapy; Mammalian Cell; Mediating; Meningoencephalitis; Modeling; Molecular; Mus; Mycoses; NIH Program Announcements; Opportunistic Infections; Patch-Clamp Techniques; Patients; Pharmaceutical Preparations; Physiological Processes; Physiology; Play; Preclinical Drug Evaluation; Role; Screening procedure; Signal Transduction; Structure of thyroid parafollicular cell; T-Lymphocyte; Testing; Tissues; Triazoles; Virulence; base; channel blockers; drug development; endoplasmic reticulum stress; high throughput screening; immune function; in vivo; inhibitor/antagonist; mortality; mutant; novel; novel strategies; patch clamp; pathogen; prevent; small molecule; small molecule libraries; synergism; therapeutic target; tool

DESCRIPTION (provided by applicant): We have developed a simple and reliable assay to identify small molecule inhibitors of the Cch1-Mid1 Ca2+ channel. This complex promotes many critical aspects of fungal physiology. In Cryptococcus neoformans, Candida albicans and C. glabrata, Cch1-Mid1 signaling plays a crucial role in tolerating and surviving ER stress conditions that are induced by inhibitors of ergosterol biosynthesis, such as triazole antifungal drugs. C. neoformans is the leading cause of fungal meningitis in AIDS patients and other patients without adequate T-cell-dependent immune functions. Small molecule modulators would provide critical tools for studying Ca2+- mediated signaling events during ER stress and might even lead the way to the development of important adjuvants that might synergize with existing antifungals and enhance the treatment of life-threatening fungal infections. However, there are currently no inhibitors of Cch1 signaling. Our assay takes advantage of the model fungal pathogen C. neoformans, which requires the Cch1-Mid1 Ca2+ channel for its survival in low Ca2+ environments. On low Ca2+ media in the presence of small molecules that block Cch1 activity and prevent Cch1-mediated signaling, the growth of C. neoformans would be arrested. Thus the assay uses a simple readout (growth versus growth arrest) to identify small molecules that inhibit the Cch1-Mid1 channel. Because the assay is reproducible and has a simple readout, this assay is highly amenable to high-throughput screening. We will use a counter screen to prioritize the lead compounds and also validate the biological relevance of each molecule through patch clamp techniques and FRET- based Ca2+ imaging. We anticipate that a screen using this assay may provide a crucial step forward in understanding the role of the Cch1-Mid1 channel in critical Ca2+-mediated signaling events and in developing a novel strategy for treating fungal infections. This project is responsive to the program announcement for the development of assays for high-throughput drug screening. PUBLIC HEALTH RELEVANCE: This study aims to understand the role of a calcium channel that governs essential physiological processes in fungal pathogens as a means to assess its viability as a target for antifungal drug development. Because fungal infections are life threatening for patients with a suppressed immune response, the development of more efficacious and better-tolerated drugs is essential.

描述（由申请人提供）：我们开发了一种简单可靠的测定方法，用于鉴定Cch 1-Mid 1 Ca 2+通道的小分子抑制剂。这种复合物促进了真菌生理学的许多关键方面。在新生隐球菌、白色念珠菌和念珠菌中，Glabrata，Cch 1-Mid 1信号传导在耐受和存活由麦角甾醇生物合成抑制剂（例如三唑类抗真菌药物）诱导的ER应激条件中起关键作用。C.在艾滋病患者和其他没有足够的T细胞依赖性免疫功能的患者中，新型脑膜炎是真菌性脑膜炎的主要原因。小分子调节剂将为研究ER应激期间Ca 2+介导的信号传导事件提供关键工具，甚至可能导致重要佐剂的开发，这些佐剂可能与现有的抗真菌药物协同作用，并增强对危及生命的真菌感染的治疗。然而，目前还没有Cch 1信号传导的抑制剂。我们的测定利用了模式真菌病原体C。neoformans，其需要Cch 1-Mid 1 Ca 2+通道用于其在低Ca 2+环境中的存活。在低Ca ~（2+）培养基中，加入阻断Cch 1活性和阻止Cch 1介导的信号转导的小分子，C.新人类会被逮捕因此，该检测使用简单的读数（生长与生长停滞）来鉴定抑制Cch 1-Mid 1通道的小分子。因为该测定是可重复的并且具有简单的读数，所以该测定非常适合于高通量筛选。我们将使用计数器筛选优先考虑先导化合物，并通过膜片钳技术和基于FRET的Ca 2+成像验证每个分子的生物学相关性。我们预计，使用这种检测方法的屏幕可能会提供一个关键的一步，在理解的Cch 1-Mid 1通道的作用，在关键的Ca 2+介导的信号转导事件，并在开发一种新的策略，用于治疗真菌感染。该项目是响应高通量药物筛选分析开发计划的公告。公共卫生关系：本研究的目的是了解钙通道的作用，管理真菌病原体的基本生理过程作为一种手段，以评估其作为抗真菌药物开发的目标的可行性。由于真菌感染对免疫反应受抑制的患者来说是威胁生命的，因此开发更有效和耐受性更好的药物至关重要。

项目成果

Flucytosine resistance in Cryptococcus gattii is indirectly mediated by the FCY2-FCY1-FUR1 pathway.

格特隐球菌中的氟胞嘧啶抗性是由 FCY2-FCY1-FUR1 途径间接介导的。

• DOI: 10.1093/mmy/myx135
• 发表时间: 2018
• 期刊: Medical mycology
• 影响因子: 2.9
• 作者: Vu,Kiem;Thompson3rd,GeorgeR;Roe,ChandlerC;Sykes,JaneE;Dreibe,ElizabethM;Lockhart,ShawnR;Meyer,Wieland;Engelthaler,DavidM;Gelli,Angie
• 通讯作者: Gelli,Angie