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

项目摘要 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2050年，老年人口将增加到1100-1600万。尽管衰老是AD最重要的风险因素，但它 目前尚不清楚在多大程度上导致与年龄相关的正常记忆缺陷的分子变化 有助于观察到AD患者的痴呆症症状。最近，我和我的同事展示了 衰老和阿尔茨海默病模型小鼠的空间记忆缺陷对应于钙依赖的破坏 海马神经元的可塑性。尽管相似，但神经元功能障碍的程度和范围 阿尔茨海默病小鼠的记忆缺陷加剧。目前的提案旨在解决是否存在“正常”衰老问题 在这些小鼠模型中，与AD相关的记忆障碍是由于共同分子的破坏造成的 途径，或由赋予相似细胞表型的不同分子改变所致。为了做某事 为此，申请人需要在蛋白质组学和基因转导方面进行额外的监督研究培训。 在初级导师安德鲁·格林博士(生理学教授兼 国家蛋白质组学研究和开发中心)和共同导师纳沙特·格尔赫斯博士 (麻省理工学院分子、细胞生物学和解剖学助理教授)这项提议的中心目标是 协助首席调查员确立其独立性，并确保获得终身教职，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 与衰老相关的痴呆症是其基础。指导阶段将为申请者提供蛋白质组学方面的培训 鉴定和定量在“正常”衰老和海马区差异表达的膜蛋白 有记忆缺陷的AD小鼠。她还将利用基于病毒的基因转导技术获得专业知识 确认几个从头开始的“命中”在记忆功能中的作用，以及确定它们在调节中的作用 固有的神经元兴奋性和突触可塑性。在这一阶段，应聘者将获得更多 应用基于病毒的基因转导技术尝试修复小鼠记忆障碍的经验 通过下调我们的先验目标TRPC3，建立“正常”衰老和AD模型。培训议程 将妇幼保健院的实验室培训、外部实验室的专业培训机会、 正规课程、蛋白质组学和AD杂志俱乐部、研讨会和教程。这种多学科的培训 将确保她有能力设计、执行、故障排除和解释多个互补的实验 分析水平。培训环境将为职业发展提供大量机会 通过国家研究报告、合作、指导学生和培训责任人 进行研究。在独立阶段，申请者将应用她最近的培训来验证 几个新靶点在记忆功能中的作用，确定靶向干扰的潜在机制/S 在细胞/突触水平上的记忆，并试图挽救“正常”衰老小鼠模型的记忆缺陷 和AD。由于AB42水平与AD相关的记忆缺陷密切相关，我们预计 成功修复记忆缺陷还可能降低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。会议的结果 拟议的研究有可能对确定这两种疾病的新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.