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: 9278064
• 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/9278064
• 关键词: 5' -AMP-activated protein kinase; Abeta synthesis; Academic Medical Centers; Achievement; Affect; Age-associated memory impairment; Alpha Cell; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease risk; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Anatomy; Animal Model; Award; Behavioral; Biochemistry; Biogenesis; Biological Assay; Biology; Biometry; Brain; Brain Diseases; Brain region; Carrier Proteins; Cell membrane; Cellular biology; Characteristics; Chemicals; Cleaved cell; Clinical Research; Clinical Trials; Collaborations; Development; Disease Progression; Environment; Enzymes; Evolution; FRAP1 gene; Faculty Workshop; Funding; Future; Future Teacher; Genes; Genetic; Genetic Screening; Goals; Grant; Hippocampus (Brain); Human; Human Amyloid Precursor Protein; 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; Pathologic; 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; Swedish mutation; Teacher Professional Development; 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; career networking; cellular targeting; chemical genetics; entorhinal cortex; gamma secretase; genetic approach; improved; inhibitor/antagonist; meetings; monoamine; mouse model; novel; peptide B; protein transport; public health relevance; responsible research conduct; secretase; skill acquisition; 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（通过应用提供）：淀粉样蛋白β-肽（Aß）的升高和积累是动物模型和患者中阿尔茨海默氏病（AD）的标志性患者特征。 Aß的产生是由两种蛋白水解酶，β位点裂解酶（BACE1）和？secressase介导的。最近的工作已经确定了与App的Bace1裂解位点相近的突变，从而使那些具有抗AD和年龄相关的认知能力下降的人。这表明BACE1介导的APP裂解是AD风险和疾病进展的第二位，并验证了介导Bace1活动进行干预的希望。为了找到BACE1的细胞调节剂，我们采用了一种化学遗传学方法来识别可以针对新型途径的化学问题，从而影响APP的BACE1裂解。用于检测特定BACE1介导的应用裂解产品的测定法分泌Appß（Sappß），用于筛选基于细胞的平台中的化学问题。鉴定出一种命中化合物，该化合物在结构上与中枢神经系统（CNS）中活性良好的药物类别相似。为了提高效率并探索结构活动关系，随后合成了化学类似物（CNS系列），这些类似物能够减少Sappß和Aß。但是，中枢神经系统系列的特定细胞靶标仍然未知。基于描述母体化合物的药物类别的文献，报告了大脑中假定靶标的区域表达和广泛的药物测试，我们确定了候选靶标，即质膜单胺转运蛋白（PMAT）。 PMAT是一种非典型的单胺转运蛋白，属于高度可吸毒的单胺转运蛋白。在啮齿动物中，PMAT在大脑中表达在广告中受影响的区域，包括内嗅皮层和海马。我们的初步研究表明，在药理学上抑制PMAT和遗传降低神经元中的PMAT表达会导致sappß和Aß水平降低。有趣的是，用PMAT抑制剂对神经元的治疗导致AMP激活蛋白激酶（AMPK）哺乳动物雷帕霉素（MTOR）信号通路中蛋白质的翻译后改变。我们建议测试PMAT在小鼠模型和人类中有助于AD的假设，并确定有助于对Sappß和Aß生物发生作用的细胞和分子机制。这些研究的成功完成将验证PMAT作为AD中的一种新型细胞靶标，该靶标有可能在药理学上利用用于AD疗法的未来开发。我的直接职业目标是研究PMAT在小鼠模型中AD发病机理中的作用及其对人类AD病理学的潜在贡献。实现这些目标将促进我的长期研究目标，即成为一个有效且独立的研究者，为广告研究领域做出重大贡献。我希望以我的生物化学和药理学背景，将我的专家扩展到广告转化研究，包括目标验证和人类病理学。通过这个培训奖，我将能够发展运输者生物学和人类的专业知识 哥伦比亚大学病理学和细胞生物学系的联席会员（Kim and Shelanski博士）和环境非常适合实现这些目标。此外，我将通过包括华盛顿大学的PMAT专家Wang博士在内，将我的科学网络扩展到哥伦比亚大学以外。哥伦比亚大学的环境与包括但不限于Drs在内的合作者富裕。 Javitch和Vonsattel支持我的科学和职业发展。为了支持独立性，我计划提交NIH的随后R01资金申请。在准备中，我将参加哥伦比亚大学欧文临床和翻译研究所，即首个R01课程。我还将通过参与哥伦比亚大学提供的正式课程来发展生物统计学和运输蛋白生物学的技能。哥伦比亚大学还提供广泛的时刻，带来专家科学家并支持讨论和合作。此外，准备未来的教师研讨会系列和学术事务学院发展局，还提供了有关职业发展技能的半手，例如赠款写作，手稿准备和演讲。我还拥有最高的资源来培训负责任的研究。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国际协会国际会议和国际阿尔茨海默氏病和帕金森氏病的国际会议，通过参加Transporter生物学和细胞信号级联的培训。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，这是科学领域成功的关键组成部分。发展为成功的独立研究者需要勤奋的研究计划和执行，卓越的培训以及个人和机构的批判性支持，所有这些都在本申请中得到了体现。提出的科学和培训部分的完成将使我成为一个高技能和公认的AD领域的贡献者。 LT还将促进随后的事业，尽管LSS正式，职业生涯的独立发展在我持续发展中，作为该领域的成功贡献者。