The role of a novel atypical monoamine transporter in Alzheimer's disease

新型非典型单胺转运蛋白在阿尔茨海默病中的作用

• 批准号: 8917841
• 负责人: Laura Beth Johnson McIntire
• 金额: $ 12.69万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8917841
• 关键词: 5' -AMP-activated protein kinase; Academic Medical Centers; Achievement; Affect; Age-associated memory impairment; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Animal Model; Award; Behavioral; Biochemistry; Biogenesis; Biological Assay; Biology; Biometry; Brain; Brain region; Carrier Proteins; Cell membrane; Cells; Cellular biology; Characteristics; Chemicals; Cleaved cell; Clinical Research; Clinical Trials; Collaborations; Development; Disease; Disease Progression; Environment; Enzymes; Evolution; Faculty; Faculty Workshop; Funding; Future; Genes; Genetic; Genetic Screening; Goals; Grant; Health; Hippocampus (Brain); Human; Human Pathology; Institutes; International; Intervention; Knowledge; Laboratories; Lead; Learning; Literature; Manuscripts; Mediating; Memory; Mentors; Molecular; Molecular Target; Mus; Mutation; Neuraxis; Neurons; New York; Parents; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Pharmacology; Phenotype; Predisposition; Preparation; Production; Proteins; Proteolysis; RNA Interference; Regulation; Reporting; Research; Research Personnel; Resistance; Resources; Rodent; Role; Science; Scientist; Seminal; Series; Signal Pathway; Signal Transduction; Site; Structure-Activity Relationship; Testing; Tg2576; Therapeutic; Tissues; Training; Transgenes; Translational Research; United States National Institutes of Health; Universities; Ursidae Family; Validation; Washington; Work; Writing; amyloid pathology; amyloid precursor protein processing; analog; base; behavior test; beta-site APP cleaving enzyme 1; brain tissue; career; career development; cellular targeting; chemical genetics; entorhinal cortex; genetic approach; improved; inhibitor/antagonist; mTOR protein; meetings; monoamine; mouse model; novel; protein transport; responsible research conduct; secretase; skills; success; symposium; translational study

DESCRIPTION (provided by applicant): Elevation and accumulation of amyloid ß-peptide (Aß) are hallmark pathological characteristics of Alzheimer's disease (AD) in animal models and patients. The production of Aß is mediated by cleavage of amyloid precursor protein (APP) by two proteolytic enzymes, ß-site cleaving enzyme (BACE1) and ?- secretase. Recent work has identified a mutation in close proximity to the BACE1 cleavage site of APP rendering those who bear the mutation resistant to AD and age-associated cognitive decline. This indicates BACE1 mediated cleavage of APP is seminal for AD risk and disease progression, and validates the promise of mediating BACE1 activity for intervention. To find cellular regulators of BACE1, we employed a chemical genetics approach to identify chemical probes that that can target novel pathways which effect BACE1 cleavage of APP. An assay for detecting the specific BACE1 mediated APP cleavage product, secreted APPß (sAPPß), was used to screen for chemical probes in a cell-based platform. A hit compound was identified which was structurally similar to a well characterized drug class active in the central nervous system (CNS). To improve the efficacy and explore the structure activity relationship, chemical analogs were subsequently synthesized (CNS-series) which were able to reduce sAPPß and Aß. However, the specific cellular target of the CNS-series remained unknown. Based on literature describing the drug class of the parent compound, the reported regional expression of putative targets in brain and extensive pharmacological testing, we identified a candidate target, the plasma membrane monoamine transporter (PMAT). PMAT is an atypical monoamine transporter belonging to a highly druggable class of monoaomine transporters. In rodents, PMAT is expressed in the brain in regions shown to be affected in AD including entorhinal cortex and hippocampus. Our preliminary studies indicated that pharmacologically inhibiting PMAT and genetically reducing PMAT expression in neurons resulted in decreased sAPPß and Aß levels. Interestingly, treatment of neurons with PMAT inhibitors led to post-translational alterations of proteins in AMP-activated protein kinase (AMPK) mammalian target of rapamycin (mTOR) signaling pathways. We propose to test the hypotheses that PMAT contributes to AD in a mouse model and in human and identify the cellular and molecular mechanisms contributing to the effect on sAPPß and Aß biogenesis. Successful completion of these studies will validate PMAT as a novel cellular target in AD which has potential to be pharmacologically harnessed for future development of AD therapeutics. My immediate career goals are to study the role of PMAT in pathogenesis of AD in a mouse model and its potential contribution to AD pathology in human. Achievement of these goals will facilitate my long-term research goal which is to become an effective and productive independent investigator to contribute significantly to the field of AD research. With my background in biochemistry and pharmacology, I hope to expand my expertise into translational studies of AD including target validation and human pathology. With this training award I will be able to develop expertise in transporter biology as well as the human pathology of AD. The co-mentors (Drs. Kim and Shelanski) and environment in the Department of Pathology and Cell Biology at Columbia University are uniquely suited for achievement of these goals. Additionally, I will expand my scientific network outside of Columbia University by including collaborator, Dr. Wang, a PMAT expert from the University of Washington. The environment at Columbia University is rich with collaborators including but certainly not limited to Drs. Javitch and Vonsattel, facilitating my scientific and career development. To facilitate independence, I plan on submitting an application for subsequent R01 funding from NIH. In preparation, I will take the Irving Institute for Clinical and Translational Research at Columbia University, Reach for the First R01 course. I also will develop skills in biostatistics and transporter biology by partaking in formal coursework offered by Columbia University. Columbia University also offers extensive seminars bringing expert scientists and facilitating discussions and collaborations. Additionally, the Preparing Future Faculty Seminar Series and Office of Academic Affairs Faculty Development Series offers seminars on career development skills such as grant writing, manuscript preparation and presentations. I also have superlative resources for training in Responsible Conduct of Research. Finally, I propose to gain training in both transporter biology and cell signaling cascades through attendance at Gordon Research Seminars and Conferences, Cold Spring Harbor Laboratory Meetings as well as international meetings in including Alzheimer's Association International Conference and International Conference on Alzheimer's disease and Parkinson's disease. Each of these opportunities will allow me to develop specific knowledge as well as my career network which are critical components for success in the field of science. Development into a successful independent investigator requires diligent research planning and execution, exceptional training and critical support, personally and institutionally, all of which are exemplified in this application. Completin of both the scientific and training portions proposed will enable my emergence as a highly skilled and recognized contributor to the field of AD. lt will also facilitate subsequent, though lss formal, career-long independent development during my continual evolution as a successful contributor to the field.

描述（由申请人提供）：淀粉样蛋白β-肽（A β）的升高和积累是动物模型和患者中阿尔茨海默病（AD）的标志性病理特征。淀粉样前体蛋白（APP）的产生是由两种蛋白水解酶，β-位点裂解酶（BACE 1）和β-淀粉样前体蛋白（APP）的裂解介导的。分泌酶最近的研究发现，APP的BACE 1切割位点附近存在一种突变，使携带该突变的人对AD和年龄相关的认知能力下降具有抵抗力。这表明BACE 1介导的APP裂解对AD风险和疾病进展至关重要，并验证了介导BACE 1活性进行干预的前景。为了找到BACE 1的细胞调节因子，我们采用化学遗传学方法来鉴定可以靶向影响BACE 1切割APP的新途径的化学探针。用于检测特异性BACE 1介导的APP切割产物分泌型APP酶（sAPP酶）的测定用于在基于细胞的平台中筛选化学探针。鉴定了一种命中化合物，其在结构上类似于在中枢神经系统（CNS）中具有活性的充分表征的药物类别。为了提高疗效和探索构效关系，随后合成了能够降低sAPP和AAPP 3的化学类似物（CNS系列）。然而，CNS系列的特异性细胞靶点仍然未知。基于描述母体化合物的药物类别的文献、报告的脑中推定靶点的区域表达和广泛的药理学测试，我们确定了一个候选靶点，即质膜单胺转运蛋白（PMAT）。PMAT是一种非典型的单胺转运蛋白，属于一类高度可药物化的单胺转运蛋白。在啮齿类动物中，PMAT在显示在AD中受影响的脑区域中表达，包括内嗅皮层和海马。我们的初步研究表明，抑制PMAT和遗传减少PMAT表达的神经元中的sAPP 3和AAPP 4水平降低。有趣的是，用PMAT抑制剂处理神经元导致AMP活化蛋白激酶（AMPK）哺乳动物雷帕霉素靶蛋白（mTOR）信号通路中蛋白质的翻译后改变。我们建议测试的假设，PMAT有助于AD在小鼠模型和人类，并确定的细胞和分子机制，有助于对sAPPPARY和AAPPPARY生物合成的影响。这些研究的成功完成将验证PMAT作为AD中的新细胞靶点，其有可能被用于AD治疗剂的未来开发。我的近期职业目标是研究PMAT在小鼠模型中AD发病机制中的作用及其对人类AD病理学的潜在贡献。这些目标的实现将有助于我的长期研究目标，即成为一名有效和富有成效的独立研究人员，为AD研究领域做出重大贡献。凭借我的生物化学和药理学背景，我希望将我的专业知识扩展到AD的转化研究，包括靶标验证和人类病理学。有了这个培训奖，我将能够发展运输生物学和人类生物学方面的专业知识 AD的病理学共同导师（金博士和Shelanski）和环境在病理学和细胞生物学系在哥伦比亚大学是唯一适合实现这些目标。此外，我将扩大我的科学网络以外的哥伦比亚大学，包括合作者，王博士，一个PMAT专家从华盛顿大学。在哥伦比亚大学的环境是丰富的合作者，包括但当然不限于博士Javitch和冯萨特尔，促进我的科学和职业发展。为了促进独立性，我计划向NIH提交后续R 01资助申请。为了准备，我将参加哥伦比亚大学欧文临床和转化研究所的第一门R 01课程。我还将通过参加哥伦比亚大学提供的正式课程来发展生物统计学和转运生物学方面的技能。哥伦比亚大学还提供广泛的研讨会，带来专家科学家和促进讨论和合作。此外，准备未来教师研讨会系列和学术事务办公室教师发展系列提供有关职业发展技能的研讨会，例如资助写作、手稿准备和演讲。我也有最好的资源来培训负责任的研究行为。最后，我建议通过参加戈登研究研讨会和会议，冷泉港实验室会议以及包括阿尔茨海默氏症协会国际会议和阿尔茨海默氏症和帕金森氏症国际会议在内的国际会议来获得转运蛋白生物学和细胞信号级联方面的培训。这些机会中的每一个都将使我能够发展特定的知识以及我的职业网络，这是在科学领域取得成功的关键组成部分。发展成为一个成功的独立调查员需要勤奋的研究规划和执行，特殊的培训和关键的支持，个人和机构，所有这些都体现在本申请。完成科学和培训部分的建议将使我成为一个高度熟练的和公认的贡献者领域的广告。它也将促进随后的，虽然lss正式的，职业生涯长期的独立发展，在我作为一个成功的贡献者不断演变的领域。