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: 8750033
• 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/8750033
• 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 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; abeta accumulation; 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; 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 β-peptide (Aβ) are hallmark pathological characteristics of Alzheimer's disease (AD) in animal models and patients. The production of Aβ is mediated by cleavage of amyloid precursor protein (APP) by two proteolytic enzymes, β-site cleaving enzyme (BACE1) and γ-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 APPβ (sAPPβ), 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 sAPPβ and Aβ. 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 sAPPβ and Aβ 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 sAPβ and Aβ 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）：β-淀粉样肽（Aβ）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。Aβ的产生是由淀粉样前体蛋白（APP）的裂解介导的。两种蛋白水解酶，β-位点裂解酶 (BACE1) 和 γ-分泌酶 最近的工作发现了与 BACE1 裂解非常接近的突变。这表明 BACE1 介导的 APP 裂解对于 AD 风险和疾病进展具有重要意义，并验证了介导 BACE1 活性用于干预的前景。针对 BACE1，我们采用化学遗传学方法来鉴定可靶向影响 BACE1 APP 裂解的新途径的化学探针。一种用于检测特定 BACE1 介导的 APP 的测定。裂解产物分泌的 APPβ (sAPPβ) 用于在基于细胞的平台中筛选化学探针，该化合物在结构上与中枢神经系统 (CNS) 中具有良好活性的药物类别相似。为了验证功效并探索结构活性关系，随后合成了能够减少 sAPPβ 和 Aβ 的化学类似物（CNS 系列），但根据描述的文献，CNS 系列的具体细胞靶点仍然未知。根据母体化合物的药物类别、所报道的脑中推定靶标的区域表达和广泛的药理学测试，我们确定了一个候选靶标，膜单胺转运蛋白（PMAT）是一种非典型单胺转运蛋白，属于高度可成药的单胺转运蛋白类别。在啮齿类动物中，PMAT 在 AD 中受影响的大脑区域中表达，包括内嗅皮层和海马体。简而言之，用 PMAT 抑制剂治疗神经元会导致 AMP 激活蛋白激酶 (AMPK) 哺乳动物雷帕霉素靶标 (mTOR) 信号通路中的蛋白质发生翻译后改变。 PMAT 在小鼠模型和人类中促进 AD，并确定了影响 sAPβ 和 Aβ 生物发生的细胞和分子机制，这些研究的成功完成将验证 PMAT 作为 AD 的新细胞靶点。 AD 有可能在药理学上用于 AD 疗法的未来发展，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 病理学的潜在贡献将有助于实现这些目标。我的长期研究目标是成为一名有效且富有成效的独立研究者，为 AD 研究领域做出重大贡献，凭借我在生物化学和药理学方面的背景，我希望将我的专业知识扩展到 AD 的转化研究，包括目标验证和人类。病理学。培训奖我将能够发展转运蛋白生物学以及人类的专业知识 哥伦比亚大学病理学和细胞生物学系的共同导师（Kim 和 Shelanski 博士）和环境非常适合实现这些目标。此外，我将通过以下方式扩展我在哥伦比亚大学之外的科学网络。哥伦比亚大学的合作者包括但不限于 Javitch 和 Vonsattel 博士，这有助于我的科学和职业发展。提交一份申请 NIH 的后续 R01 资助。在准备过程中，我将参加哥伦比亚大学欧文临床和转化研究所的第一个 R01 课程，我还将通过参加 NIH 提供的正式课程来培养生物统计学和转运生物学的技能。哥伦比亚大学还举办研讨会，邀请专家科学家并促进讨论和合作。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广泛研讨会，例如资助写作、手稿。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。包括阿尔茨海默病协会国际会议和阿尔茨海默病和帕金森病国际会议，这些机会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。一名成功的独立研究者需要勤奋的研究规划和执行、卓越的培训和关键的个人和机构支持，所有这些都在本申请中得到了例证。完成所提出的科学和培训部分将使我成为一名高技能和公认的贡献者。在我作为该领域的成功贡献者不断发展的过程中，它也将促进后续的、虽然是正式的、职业生涯的独立发展。