Mechanisms of Dantrolene Neuroprotection in Alzheimer's Disease

丹曲林对阿尔茨海默病的神经保护机制

• 批准号: 10343664
• 负责人: HUAFENG WEI
• 金额: $ 40.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10343664
• 关键词: AD transgenic mice; Address; Affect; Agonist; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer’s disease biomarker; Amyloid; Amyloid beta-42; Animal Model; Autophagocytosis; Biological Markers; Brain; Calcium; Cell Proliferation; Cell model; Cell physiology; Cells; Clinical; Clinical Treatment; Clinical Trials; Cognition; Cytosol; Dantrolene; Data; Dementia; Development; Disease; Endoplasmic Reticulum; Fibroblasts; Functional disorder; Glutamates; Glycine Receptors; Goals; Hippocampus (Brain); Homeostasis; Human; Impaired cognition; Impairment; Inflammatory; Intranasal Administration; Memory Loss; Messenger RNA; Mitochondria; Mus; Mutation; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neuroglia; Neuronal Differentiation; Neurons; Oral; Pathologic; Pathology; Patients; Persons; Pharmaceutical Preparations; Plasma; Proteins; Risk Factors; Rodent; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Site; Skin; Synapses; Testing; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Treatment Efficacy; adult neurogenesis; amyloid pathology; antagonist; apolipoprotein E-4; cognitive function; cytokine; dentate gyrus; efficacy evaluation; experimental study; familial Alzheimer disease; improved; induced pluripotent stem cell; migration; mouse model; nerve stem cell; neurogenesis; neuropathology; neuroprotection; presenilin-1; receptor; receptor expression; side effect; stem cells; synaptogenesis

Abstract Clinical trials for the treatment of Alzheimer's disease (AD) targeting amyloid have largely failed. Ca2+ dysregulation may be an alternative mechanism. In familial AD (FAD), calcium homeostasis is disrupted by over activation of NMDA (NMDAR) and ryanodine (RYR) receptors leading to increased cytosolic calcium and subsequent cognitive dysfunction and neuropathology. APOE4, a major risk factor for sporadic AD (SAD), also causes Ca2+ dysregulation related to NMDAR/RYR over activation. Dantrolene, a RYR antagonist and clinically available drug, has been shown to mitigate amyloid pathology, neurodegeneration, synaptic and memory loss in a FAD animal model. Our preliminary studies suggested that intranasal dantrolene administration abolished memory loss in 5XFAD mice. Furthermore, dantrolene promoted neuronal differentiation in induced pluripotent stem cells (iPSC) from patients with either SAD with APOE4 or familial AD (FAD) by inhibition of RYR/NMDAR over- activation. Our long term goal is to examine the efficacy of dantrolene to treat AD. The overall objective of this study is to investigate the effects and underlying mechanisms of dantrolene on adult neurogenesis in human and rodent SAD and FAD cells, as well as the effects of intranasal dantrolene administration on adult neurogenesis and cognitive function in AD transgenic mice. Our central hypothesis is that intranasal dantrolene inhibits the excessive activation of RYRs and NMDARs in AD and promotes adult neurogenesis, improved cognitive function and reduced AD neuropathology. We will test the hypothesis by the following specific aims. Specific Aim 1. To determine the effects of dantrolene on RYR and NMDAR expression and on cytosolic, mitochondrial, and ER Ca2+ concentrations and AD biomarkers in AD stem cells using induced pluripotent stem cells (iPSC) from fibroblasts patients with either SAD (APOE4 risk factor) or FAD (PSEN1 mutations), as well as neuroprogenitor cells (NPC) isolated from E4FAD (APOE4+5XFAD) and 5XFAD transgenic Specific Aim 2. To examine the effects of dantrolene on RYR and NMDAR activity, neurogenesis, proliferation, and the cellular function of derived neurons and glia from AD cells. using the same cells as in SA1. Specific Aim 3. To determine the effects of dantrolene on adult neurogenesis, cognitive function, and neuropathology in animal models of AD. We expect that dantrolene will promote adult neurogenesis and restore cognition and reduce AD pathology in AD mouse models by alleviating the excessive activation of RYRs and/or NMDARs, especially with the intranasal approach.

抽象的 针对淀粉样蛋白治疗阿尔茨海默病 (AD) 的临床试验 很大程度上失败了。 Ca2+ 失调可能是另一种机制。在家族性AD（FAD）中， NMDA (NMDAR) 和兰尼定 (RYR) 的过度激活会破坏钙稳态 受体导致胞质钙增加和随后的认知功能障碍 神经病理学。 APOE4 是散发性 AD (SAD) 的主要危险因素，也会导致 Ca2+ 与 NMDAR/RYR 过度激活相关的失调。 Dantrolene，一种 RYR 拮抗剂 临床上可用的药物，已被证明可以减轻淀粉样蛋白病理、神经变性、 FAD 动物模型中的突触和记忆丧失。我们的初步研究表明 鼻内丹曲林给药消除了 5XFAD 小鼠的记忆丧失。此外， 丹曲林促进诱导多能干细胞 (iPSC) 的神经元分化 通过过度抑制 RYR/NMDAR 来治疗患有 APOE4 的 SAD 或家族性 AD (FAD) 的患者 激活。我们的长期目标是检查丹曲林治疗 AD 的功效。整体 本研究的目的是调查丹曲林对 人类和啮齿动物 SAD 和 FAD 细胞的成体神经发生，以及鼻内 丹曲林给药对 AD 转基因小鼠成年神经发生和认知功能的影响。 我们的中心假设是鼻内丹曲林抑制过度激活 RYR 和 NMDAR 在 AD 中促进成人神经发生，改善认知功能 并减少 AD 神经病理学。我们将通过以下具体目标来检验该假设。 具体目标 1. 确定丹曲林对 RYR 和 NMDAR 表达的影响 以及 AD 中的细胞质、线粒体和 ER Ca2+ 浓度以及 AD 生物标志物 使用来自患有 SAD 的成纤维细胞的诱导多能干细胞 (iPSC) 的干细胞 （APOE4 危险因素）或 FAD（PSEN1 突变），以及分离的神经祖细胞 (NPC) 来自 E4FAD (APOE4+5XFAD) 和 5XFAD 转基因 具体目标 2. 检查效果 丹曲林对 RYR 和 NMDAR 活性、神经发生、增殖和细胞 AD 细胞衍生的神经元和神经胶质细胞的功能。使用与 SA1 中相同的单元格。 具体目标 3. 确定丹曲林对成人神经发生、认知能力的影响 AD 动物模型中的功能和神经病理学。我们预计丹曲林将 促进 AD 小鼠的成年神经发生并恢复认知并减少 AD 病理 通过减轻 RYR 和/或 NMDAR 的过度激活来建立模型，特别是 鼻内方法。