Proteomics of memory failure: unraveling the relationship between 'normal' brain

项目摘要 阿尔茨海默氏病（AD）痴呆目前在美国折磨着超过500万人，并且预计 到2050年将增加到1100万至1600万老年人。虽然衰老是AD最重要的危险因素，但它 目前还不清楚在多大程度上，“正常”的年龄相关的记忆缺陷的分子变化， 有助于在AD患者中观察到的痴呆症状。最近我和我的同事证明了 衰老和AD小鼠模型中的空间记忆缺陷对应于Ca 2+依赖的 海马神经元的可塑性。尽管类似，神经元功能障碍的程度和范围， 记忆缺陷在AD小鼠中加重。目前的建议旨在解决“正常”老龄化是否 在这些小鼠模型中，AD相关的记忆障碍是由共同的分子破坏引起的。 途径，或由赋予相似细胞表型的不同分子改变引起。为做 因此，申请人需要在蛋白质组学和基因转导方面进行额外的监督研究培训 安德鲁格林博士（生理学教授和主任的主要导师的指导下， MCW国家蛋白质组学研究与开发中心）和共同导师Nashaat Gerges博士 （助理教授，分子，细胞生物学和解剖学在MCW）。该提案的核心目标是 协助首席研究员建立她的独立性，并确保终身教职，如 她可以领导一个主要的研究项目，旨在确定易感性和因果因素， 与衰老有关的痴呆症指导阶段将为申请人提供蛋白质组学培训， 鉴定和定量在“正常”老化海马中差异表达的膜蛋白， AD小鼠的记忆缺陷。她还将获得使用基于病毒的基因转导技术的专业知识， 验证几个从头'命中'在记忆功能中的作用，以及确定它们在调节记忆功能中的作用。 内在神经元兴奋性和突触可塑性。在这个阶段，候选人将获得更多 使用基于病毒的基因转导技术尝试挽救小鼠记忆缺陷的经验 通过下调我们的先验靶点TRPC 3来研究“正常”衰老和AD模型。培训议程 结合了MCW的实验室培训，外部实验室的专业培训机会， 正式的课程，蛋白质组学和AD期刊俱乐部，研讨会和教程。这种多学科培训 将确保她的能力，设计，执行，故障排除和解释实验在多个，互补 分析水平。培训环境将为职业发展提供众多机会 通过国家研究报告，合作，指导学生，并对负责任的 进行研究。在独立阶段，申请人将应用其最近的培训来验证 几个新的目标在记忆功能中的作用，确定了潜在的靶向破坏的机制， 在细胞/突触水平的记忆，并试图挽救记忆缺陷的小鼠模型的“正常”老化 的AD。由于Ab 42水平与AD相关的记忆缺陷密切相关2，我们预计， 成功挽救记忆缺陷也可能降低Ab 42水平，这将与 西北大学医学院的阿尔茨海默病专家罗伯特·瓦萨博士说。成果 拟议的研究有可能对确定新的治疗方法产生重大影响， 衰老和AD相关的记忆障碍。

Project Summary Alzheimer's Disease (AD) dementia currently afflicts over 5 million people in the United States and is projected to rise to 11-16 million elderly by the year 2050. Although aging is the most important risk factor for AD, it remains unclear to what extent the molecular changes that underlie 'normal' age-associated memory deficits contribute to symptoms of dementia observed in patients with AD. Recently my colleagues and I demonstrated that spatial memory deficits in mouse models of aging and AD correspond to disruption of Ca2+-dependent plasticity in neurons of the hippocampus. Albeit similar, the magnitude of neuronal dysfunction and scope of memory deficits were exacerbated in AD mice. The present proposal seeks to address whether 'normal' aging and AD-related memory impairments in these mouse models result from disruption of common molecular pathways, or result from divergent molecular alterations that confer similar cellular phenotypes. In order to do this, the applicant requires additional supervised research training in proteomics and gene transduction systems under the direction of primary mentor Dr. Andrew Greene (Professor of Physiology and Director of a National Center for Proteomics Research and Development at MCW) and co-mentor Dr. Nashaat Gerges (Assistant Professor, Molecular, Cell Biology and Anatomy at MCW). The central goal of this proposal is to assist the Principal Investigator establish her independence and secure a tenure-track faculty position such that she can lead a major research program aimed at determining susceptibility and causal factors that underlie aging-related dementias. The Mentored Phase will provide the applicant with training in proteomics to identify and quantitate membrane proteins differentially expressed in the hippocampus of 'normal' aging and AD mice with memory deficits. She will also gain expertise using viral-based gene transduction techniques to validate the role of several de novo 'hits' in memory function, as well as determine their role in modulating intrinsic neuronal excitability and synaptic plasticity. During this phase, the candidate will gain further experience using viral-based gene transduction techniques to attempt the rescue of memory deficits in mouse models of 'normal' aging and AD by downregulating our a priori target TRPC3. The training agenda incorporates laboratory-based training at MCW, opportunities for specialty training in external laboratories, formal coursework, proteomics and AD journal clubs, seminars, and tutorials. Such multidisciplinary training will ensure her ability to design, perform, troubleshoot and interpret experiments at multiple, complementary levels of analysis. The training environment will provide numerous opportunities for career development through national research presentations, collaborations, mentoring students, and training on the responsible conduct of research. During the Independent phase, the applicant will apply her recent training to validate the role of several novel targets in memory function, determine the mechanism/s underlying targeted disruption of memory at the cellular/synaptic level, and attempt to rescue memory deficits in mouse models of 'normal' aging and AD. Because Ab42 levels are strongly correlated with AD-related memory deficits2, we expect that successful rescue of memory deficits may also reduce Ab42 levels that will be tested in collaboration with Alzheimer's Disease expert Dr. Robert Vassar at Northwestern University Medical School. Outcomes of the proposed research have the potential to make a major impact on the identification of new treatments for both aging and AD-related memory disorders.