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

描述（由申请人提供）：阿尔茨海默病（AD）痴呆目前在美国折磨着超过500万人，预计到2010年将上升到1100万至1600万老年人。 2050年。虽然衰老是AD最重要的危险因素，但目前尚不清楚在AD患者中观察到的痴呆症状中，“正常”年龄相关记忆缺陷的分子变化在多大程度上起作用。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与海马神经元中Ca2+依赖的可塑性的破坏相对应。尽管相似，但AD小鼠的神经元功能障碍程度和记忆缺陷范围均加重。本提案旨在解决这些小鼠模型中的"正常"衰老和AD相关记忆障碍是否是由于常见分子途径的破坏，或者是由于赋予相似细胞表型的不同分子改变。为做 因此，申请人需要在主要导师Andrew格林博士（MCW的生理学教授和国家蛋白质组学研究与开发中心主任）和共同导师Nashaat Gerges博士（MCW的分子、细胞生物学和解剖学助理教授）的指导下进行蛋白质组学和基因转导系统的额外监督研究培训。该提案的中心目标是帮助主要研究者建立她的独立性，并确保终身教职，使她能够领导一个主要的研究计划，旨在确定易感性和因果因素，这些因素是与衰老相关的痴呆症的基础。指导阶段将为申请人提供蛋白质组学方面的培训，以鉴定和定量在"正常"衰老和记忆缺陷的AD小鼠的海马体中差异表达的膜蛋白。她还将获得使用基于病毒的基因转导技术的专业知识，以验证几个从头“命中”在记忆功能中的作用，以及确定它们在调节内在神经元兴奋性和突触可塑性中的作用。在此阶段，候选人将获得进一步的经验，使用基于病毒的基因转导技术，试图通过下调我们的先验靶点TRPC 3来挽救"正常"衰老和AD小鼠模型中的记忆缺陷。培训议程包括MCW的实验室培训，外部实验室专业培训的机会，正式课程，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有能力设计，执行，故障排除和解释实验在多个，互补的分析水平。培训环境将通过国家研究报告，合作，指导学生和负责任的研究行为的培训，为职业发展提供许多机会。在独立阶段，申请人将应用她最近的培训，以验证几个角色 新的目标在记忆功能，确定机制/s潜在的有针对性的破坏记忆在细胞/突触水平，并试图挽救记忆缺陷的小鼠模型的"正常"老化和AD。由于Ab42水平与AD相关的记忆缺陷密切相关，我们预计成功挽救记忆缺陷也可能降低Ab42水平，这将与西北大学医学院的阿尔茨海默病专家Robert Vassar博士合作进行测试。拟议研究的结果有可能对识别衰老和AD相关记忆障碍的新疗法产生重大影响。 公共卫生相关性：该项目的意义在于，它将采用一种新的方法来识别导致“正常”衰老和阿尔茨海默病中发生的记忆缺陷的新分子，并使用基因治疗方法来治疗衰老和阿尔茨海默病小鼠模型的记忆障碍。拟议研究的结果有很大的潜力，对确定与年龄相关的认知能力下降和阿尔茨海默病痴呆症的新治疗方法产生重大影响。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The significance of this project is that it will employ a novel approach to identify new molecules that contribute to memory deficits that occur in 'normal' aging and Alzheimer's Disease, and use gene therapy approaches to treat memory failure in mouse models of aging and Alzheimer's Disease. Outcomes of the proposed research have great potential to make a major impact on the identification of new treatments for both age-associated cognitive decline and Alzheimer's Disease dementia.