Restorative gene therapy on hippocampal spino-dendritic abnormalities: a focus on Alzheimer’s disease

海马棘树突异常的恢复性基因治疗：关注阿尔茨海默病

• 批准号: 10343842
• 负责人: RUTH M BARRIENTOS
• 金额: $ 19.83万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10343842
• 关键词: 3xTg-AD mouse; Adult; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Behavioral; Behavioral Paradigm; Brain region; Candidate Disease Gene; Characteristics; Clinical; Clinical Trials; Confocal Microscopy; Data; Dementia; Dendrite Regeneration; Dendrites; Dendritic Spines; Disease; Elderly; Exhibits; Exposure to; Female; Genes; Goals; Golgi Apparatus; Hippocampus (Brain); Immunohistochemistry; Impaired cognition; Impairment; In Vitro; Injections; Intracellular Signaling Proteins; Knowledge; Laboratories; Learning; Locomotion; Long-Term Potentiation; Maintenance; Measurement; Measures; Mediating; Mediator of activation protein; Memory; Memory Loss; Memory impairment; Mission; Morphogenesis; Morphology; Motor; Mouse Strains; Mus; Mutation; Natural regeneration; Neurites; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Neurotoxins; Pathologic; Pediatric Hospitals; Pharmaceutical Preparations; Phosphoproteins; Process; Proteins; Public Health; Quality of life; Research; Role; Semaphorin-3A; Senile Plaques; Serotyping; Stains; Structure; Synapses; Synaptic plasticity; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Vertebral column; Viral; Wild Type Mouse; Work; adeno-associated viral vector; cognitive function; conditioned fear; density; design; effective intervention; effective therapy; experience; gene therapy; generalized anxiety; healthy aging; improved; in vivo; innovation; male; morris water maze; mouse model; neuropathology; novel; overexpression; presenilin-1; preservation; prevent; repaired; response; spatial memory; therapeutically effective; vector

SUMMARY/ABSTRACT There are currently no effective treatments to prevent, delay, or reverse the memory deficits and neuropathological alterations—specifically dendritic damage and synaptic loss—associated with Alzheimer’s disease (AD). Without an effective treatment soon, many more millions of older adults will continue to suffer from this devastating disease. Thus, there is a critical need to investigate novel molecules/mechanisms involved in the maintenance and regeneration of dendrites and their spines for the purpose of rescuing cognitive functions. The long-term goal is to develop an effective treatment to prevent or halt AD. The overall objective of this application is to determine the potential of a recently identified phosphoprotein, CRMP3 (collapsing response mediator protein 3), to ameliorate dendritic abnormalities and/or rescue memory deficits in two distinct mouse models with AD-like characteristics. 3xTg-AD mice are one of the most commonly used mouse models of AD. They harbor 3 mutations (APP, PSEN1, and MAPT) that result in the accumulation of plaques and tangles, abnormalities in dendritic and synaptic structures, and significant cognitive impairments. CRMP3-/- mice exhibit dystrophic hippocampal dendrites and profoundly impaired synaptic plasticity (LTP). Importantly, overexpressing the CRMP3 protein in hippocampal neurons, in vitro, dramatically enhances dendritic formation. The central hypothesis is that over-expressing CRMP3, in vivo, will prevent further dendritic degeneration and/or evoke dendritic regrowth and remodeling of spines in otherwise degenerating hippocampal neurons, thus preserving or rescuing memory function. This hypothesis has been formulated on the basis of strong preliminary data produced in the applicants’ laboratories. This hypothesis will be tested by pursuing two specific aims: 1) Determine the extent to which in vivo over-expression of CRMP3 enhances hippocampal dendritic complexity and spine structure in CRMP3-/- and 3xTg-AD mice; and 2) Determine the extent to which in vivo over-expression of CRMP3 affects hippocampal-dependent memory function in CRMP3-/- and 3xTg-AD mice. Under the first aim, adeno-associated viral vector serotype 9 (AAV9) gene therapy will be used to overexpress CRMP3 in the hippocampus of these two AD-like mouse models and its effects on AD-associated spino-dendritic abnormalities will be examined. Under the second aim, the extent to which CRMP3 gene therapy can ameliorate contextual and spatial memory function will be measured using two behavioral paradigms (contextual fear conditioning and Morris water maze). The approach is innovative because CRMP3 is a little-known protein with tremendous potential to repair and regenerate dystrophic and damaged spines and dendrites. The proposed research is significant because it is expected to advance knowledge about a protein with the robust capacity to slow, halt, or even reverse early AD processes that may be precursors to deep dementia, thus profoundly improving quality of life. Ultimately, such findings have the potential to move to clinical trials as a treatment for AD.

摘要/摘要 目前尚无有效的治疗方法来预防、延缓或逆转记忆缺陷和 与阿尔茨海默病相关的神经病理学改变，特别是树突损伤和突触丧失 疾病（AD）。如果不能尽快得到有效的治疗，数以百万计的老年人将继续遭受这种疾病的折磨 这种毁灭性的疾病。因此，迫切需要研究涉及的新分子/机制 树突及其脊柱的维持和再生，以挽救认知功能。 长期目标是开发一种有效的治疗方法来预防或阻止 AD。本次活动的总体目标 该应用的目的是确定最近发现的磷蛋白 CRMP3（崩溃反应）的潜力 介导蛋白 3)，改善两种不同小鼠的树突异常和/或挽救记忆缺陷 具有类似 AD 特征的模型。 3xTg-AD 小鼠是最常用的 AD 小鼠模型之一。 它们含有 3 种突变（APP、PSEN1 和 MAPT），导致斑块和缠结的积累， 树突和突触结构异常，以及严重的认知障碍。 CRMP3-/- 小鼠表现出 营养不良的海马树突和严重受损的突触可塑性（LTP）。重要的是，过度表达 在体外，海马神经元中的 CRMP3 蛋白可显着增强树突的形成。中央 假设体内过度表达 CRMP3 将阻止进一步的树突变性和/或诱发 dendritic regrowth and remodeling of spines in otherwise degenerating hippocampal neurons, thus preserving 或拯救记忆功能。这一假设是在强有力的初步数据的基础上提出的 在申请人的实验室生产。该假设将通过追求两个具体目标来检验：1） 确定 CRMP3 体内过度表达增强海马树突复杂性的程度 CRMP3-/- 和 3xTg-AD 小鼠的脊柱结构； 2) 确定体内过度表达的程度 CRMP3 影响 CRMP3-/- 和 3xTg-AD 小鼠的海马依赖性记忆功能。在第一个目标下， 腺相关病毒载体血清型 9 (AAV9) 基因治疗将用于在 这两种 AD 样小鼠模型的海马及其对 AD 相关棘树突异常的影响 将接受检查。在第二个目标下，CRMP3基因治疗可以在多大程度上改善背景 空间记忆功能将使用两种行为范式（情境恐惧调节和 莫里斯水迷宫）。该方法具有创新性，因为 CRMP3 是一种鲜为人知的蛋白质，具有巨大的作用。 修复和再生营养不良和受损的棘和树突的潜力。拟议的研究是 意义重大，因为它有望推进有关蛋白质的知识，该蛋白质具有减缓、停止、 甚至逆转可能是深度痴呆症前兆的早期 AD 过程，从而显着提高质量 的生活。最终，这些发现有可能进入临床试验，作为 AD 的治疗方法。