NMDA receptor trafficking by the autophagy regulatory protein beclin 1

描述(申请人提供)：这份K08指导临床医生-科学家职业发展奖申请书，题为《自噬调控蛋白Beclin 1的NMDA受体贩运》，由斯坦福大学医学院病理学助理教授Edward D.Plowey医学博士提交。普洛维博士是一名神经病理学家和临床科学家，他的研究兴趣在于神经退行性变，以及突触和神经元应对或屈服于神经退行性疾病应激源的机制。普洛维博士目前的研究目标是调查自噬在调节神经退行性疾病应激源的适应性突触调节中扮演的新角色，这是他独立研究计划的基石。在这项研究中，提出了一种新的机制，即依赖于Beclin的自噬来调节突触与突触外NMDA受体的定位、亚型、内化和降解。他将验证他的假设，即依赖于Beclin的自噬介导了NMDA受体池的神经保护性调节。在本研究方案的目标1中，将研究突触和突触外NMDA受体激活对Beclin 1水平、BECN1基因表达和神经保护性Beclin依赖的自噬能力的影响。在第二个目标中，将描述贝克林缺乏对表面NMDA受体水平、定位、亚型分布和毒性的影响。在目标3中，分子 在此基础上，将研究APP对Beclin-1与NMDA受体相互作用的转运效应和调节作用，重点是Beclin-1通过掺入质膜来源的自噬前体小泡而靶向表面NR1/NR2B受体降解自噬小体的可能性。普洛维博士将了解自噬如何调节神经元蛋白质的周转和突触干扰后的兴奋性，以及依赖于Beclin的自噬如何影响NMDA受体的运输以及突触和突触外NMDA受体的平衡。此外，他将拓宽我们对依赖于Beclin的自噬作为神经退行性变治疗靶点的潜在知识。Plowey博士的职业发展计划包括在突触神经生物学技术方面的高级研究培训，包括受体运输研究、膜片钳电生理学、器官型切片培养、亚细胞分离方法、分子相互作用研究以及研究突触和突触外NMDA受体的定位和功能输出的方法。他还将参加培训活动，通过专业服务、讲习班和会议提高他的赠款写作和手稿写作技能，并扩大他的专业网络和在科学界的知名度。斯坦福大学的大学学术环境是临床医生-科学家事业成功发展的理想选择。作为斯坦福大学医学院病理学系的一名独立临床医生兼科学家，Plowey博士对他的职业发展有着坚定的制度承诺，包括承诺实验室空间、慷慨的实验室启动资金和75%的受保护时间用于他的研究计划和职业发展活动。此外，Plowey博士还与在Tony Wyss-Coray博士和Craig Garner博士中拥有丰富指导经验的富有成效的高级研究人员建立了理想的导师-门生关系。随着K08职业发展奖支持他的研究和职业发展计划，Plowey博士将作为一名执业神经病理学家和NIH R01资助的神经变性突触神经生物学可持续研究项目的首席研究员，为他的长期目标做好准备。

DESCRIPTION (provided by applicant): This K08 Mentored Clinician-Scientist Career Development Award application, entitled "NMDA Receptor Trafficking by the Autophagy Regulatory Protein Beclin 1", is submitted by Edward D. Plowey, M.D., Ph.D., Assistant Professor of Pathology at Stanford University School of Medicine. Dr. Plowey is a neuropathologist and clinician-scientist whose research interests lie in neurodegeneration and the mechanisms through which synapses and neurons cope with or succumb to neurodegenerative disease stressors. Dr. Plowey's immediate research goal is to investigate novel roles for autophagy in mediating adaptive synaptic adjustments to neurodegenerative disease stressors as the cornerstone of his independent research program. In this research proposal, a focus on beclin-dependent autophagy as a novel mechanism to regulate synaptic versus extra synaptic localization, subtype, internalization and degradation of NMDA receptors is proposed. He will test his hypothesis that beclin-dependent autophagy mediates neuroprotective modulation of NMDA receptor pools. In Aim 1 of this research proposal, the effects of synaptic versus extra synaptic NMDA receptor activation on beclin 1 levels, BECN1 gene expression and the capacity for neuroprotective beclin-dependent autophagy will be investigated. In the second Aim, the impact of beclin-deficiency on surface NMDA receptor levels, localization, subtype distribution and toxicity will be delineated. In Aim 3, the molecular basis, trafficking effects and modulatory effects of APP on beclin 1 interactions with NMDA receptors will be investigated, with a focus on the possibility that beclin 1 targets surface NR1/NR2B receptors for autophagolysosomal degradation by incorporation into plasma membrane derived autophagosomal precursor vesicles. Dr. Plowey will learn how autophagy modulates neuronal protein turnover and excitability secondary to synaptic perturbations and how beclin-dependent autophagy impacts NMDA receptor trafficking and the balance of synaptic and extra synaptic NMDA receptors. Furthermore, he will broaden our knowledge of the potential for beclin-dependent autophagy as a therapeutic target in neurodegeneration. Dr. Plowey's career development plan includes advanced research training in synaptic neurobiology techniques including receptor trafficking studies, patch clamp electrophysiology, organotypic slice cultures, subcellular fractionation approaches, molecular interaction studies and approaches to study the localization and functional outputs of synaptic and extra synaptic NMDA receptors. He will also engage in training activities to improve his grant writing and manuscript writing skills and increase his professional network and visibility in the scientific community through professional service, workshops and conferences. The collegial academic environment at Stanford University is ideal for the successful career development of clinician-scientists. Dr. Plowey has strong institutional commitment to his career development as an independent clinician-scientist from the Department of Pathology at Stanford University School of Medicine, which includes commitment of laboratory space, generous laboratory startup funds and 75% protected time to devote to his research program and career development activities. Furthermore, Dr. Plowey has established ideal mentor-prot¿g¿ relationships with productive senior investigators with extensive mentorship experience in Tony Wyss-Coray, Ph.D. and Craig Garner, Ph.D. With a K08 Career Development Award to support his research and career development plans, Dr. Plowey will be positioned for success in his long term goals as a practicing neuropathologist and as a principal investigator of a sustainable NIH R01 funded research program in the synaptic neurobiology of neurodegeneration.