Proteomics of memory: Normal brain aging and Alzheimers Disease

描述(申请人提供)：阿尔茨海默病(AD)痴呆症目前在美国困扰着500多万人，预计到2019年将增加到1100-1600万老年人 公元2050年。尽管衰老是阿尔茨海默病最重要的风险因素，但目前尚不清楚与年龄相关的正常记忆缺陷背后的分子变化在多大程度上导致了阿尔茨海默病患者的痴呆症状。最近，我和我的同事证明，衰老和阿尔茨海默病小鼠模型中的空间记忆缺陷对应于海马神经元钙依赖可塑性的破坏。尽管相似，但阿尔茨海默病小鼠的神经元功能障碍的程度和记忆障碍的范围都加剧了。目前的建议试图解决这些小鼠模型中“正常”衰老和AD相关的记忆障碍是由于共同的分子通路被破坏，还是由于具有相似细胞表型的不同分子改变所致。为了做某事 为此，申请人需要在主要导师Andrew Greene博士(生理学教授和MCW国家蛋白质组学研究和开发中心主任)和共同导师Nashaat Gerges博士(MCW分子、细胞生物学和解剖学助理教授)的指导下，在蛋白质组学和基因转导系统方面进行额外的监督研究培训。这项提议的中心目标是帮助首席调查员建立她的独立性，并获得一个终身教职跟踪的教员职位，这样她就可以领导一个旨在确定与衰老相关的痴呆的易感性和因果因素的主要研究计划。指导阶段将为申请者提供蛋白质组学方面的培训，以识别和定量在“正常”衰老和记忆缺陷的AD小鼠的海马区差异表达的膜蛋白。她还将获得使用基于病毒的基因转导技术的专业知识，以验证几个从头开始的“命中”在记忆功能中的作用，以及确定它们在调节固有的神经元兴奋性和突触可塑性方面的作用。在这一阶段，候选人将获得进一步的经验，使用病毒为基础的基因转导技术，试图通过下调我们的先验目标TRPC3，来挽救‘正常’衰老和AD小鼠模型的记忆缺陷。培训议程包括MCW的实验室培训、外部实验室的专业培训机会、正式课程、蛋白质组学和AD杂志俱乐部、研讨会和教程。这样的多学科培训将确保她在多个互补的分析水平上设计、执行、故障排除和解释实验的能力。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培训，为职业发展提供大量机会。在独立阶段，申请者将应用她最近的培训来验证以下几个角色 记忆功能的新靶点，在细胞/突触水平上确定记忆靶向破坏的潜在机制/S，并试图挽救‘正常’衰老和AD模型的记忆缺陷。由于AB42水平与AD相关的记忆缺陷密切相关，我们预计，成功修复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氏症专家罗伯特·瓦萨尔博士合作进行测试。拟议的研究结果有可能对确定衰老和AD相关记忆障碍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.