Molecular mechanisms underlying heme transport at the blood-brain barrier and its role in angiogenesis

血红素在血脑屏障转运的分子机制及其在血管生成中的作用

• 批准号: 10572752
• 负责人: Rosemary Jane Cater
• 金额: $ 16.14万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10572752
• 关键词: Address; Affinity; Award; Binding; Biochemical; Biological; Biological Assay; Biological Process; Biology; Biophysics; Blood - brain barrier anatomy; Blood Vessels; Blood brain barrier dysfunction; Brain; Brain Vascular Malformation; Buffers; Carrier Proteins; Catalysis; Cell Line; Cell Surface Receptors; Cellular biology; Central Nervous System; Cerebrovascular system; Cerebrum; Co-Immunoprecipitations; Communication; Communities; Complex; Cryoelectron Microscopy; Data; Development; Development Plans; Disease; Drug Delivery Systems; Educational process of instructing; Electron Transport; Electrophysiology (science); Endothelial Cells; Environment; Fab Immunoglobulins; Faculty; Feline Leukemia Virus; Gases; Genetic; Goals; Grant; Growth; Heme; Home; Homeostasis; Human; Hydranencephaly; Hydrocephalus; Hydrophobicity; Immunoglobulin Fragments; In Vitro; Ion Channel; Job Application; Label; Laboratories; Life; Link; Lipids; Liposomes; Maps; Mediating; Membrane Proteins; Mentors; Methods; Molecular; Molecular Conformation; Mus; Mutagenesis; Mutation; Nutrient; Online Mendelian Inheritance In Man; Oxygen; Pathology; Phase; Phenotype; Phosphotransferases; Play; Positioning Attribute; Principal Investigator; Process; Professional Competence; Proliferating; Property; Protein Biochemistry; Recombinants; Research; Research Personnel; Role; Rosemary; Science; Signaling Molecule; Specificity; Stroke; Structural Biologist; Structure; Subgroup; System; Technology; Testing; Toxin; Training; Transcriptional Regulation; Transport Process; Universities; Vascular Diseases; Writing; angiogenesis; blood vessel development; brain endothelial cell; career; career development; cerebrovascular; cofactor; cytotoxic; driving force; experimental study; insight; loss of function mutation; malformation; multidisciplinary; nanodisk; nervous system development; nervous system disorder; oxygen transport; particle; programs; receptor; reconstitution; research and development; responsible research conduct; skills; structural biology; tenure track; transcription factor; uptake

PROJECT SUMMARY This application details career development and research plans that have been uniquely tailored to facilitate transition of the principal investigator, Dr. Rosemary J. Cater, to an independent academic position. Dr. Cater has a multidisciplinary background, and through this K99/R00 proposal, seeks to finalize her biochemical training, master the biology of a new system, and gain skills for her career as an independent investigator. The K99 phase of this award (first two years) involves a structured career development plan that will allow Dr. Cater to: advance her skills in single particle cryo-electron microscopy (cryo-EM); gain essential training in brain endothelial cell-based assays and proximity-based labeling experiments; and acquire important career skills such as grant writing, science communication, laboratory management, teaching, responsible conduct of research, and mastery of the academic job application process. The career development plan also includes clear and actionable steps for identifying and successfully obtaining an independent tenure-track faculty position by the end of the K99 phase. Dr. Cater has assembled a top-tier team of multi-disciplinary mentors, advisors, and collaborators that will oversee and guide her training, research program, and transition to independence. The research plan proposed spans both the mentored (K99) and independent (R00) phases of the award. It involves mechanistic studies of the heme transporter FLVCR2, which is expressed within blood-brain barrier endothelial cells and plays a key role in the development of brain vasculature. The research program for the K99 phase aims to characterize the structure and function of FLVCR2 at a molecular level. The R00 research program then aims to delineate how FLVCR2 regulates angiogenesis at a cellular level. Core research questions addressed include: What are the molecular determinants important for FLVCR2 to bind heme specifically? What conformational changes must FLVCR2 undertake to transport heme? What drives this transport process? And ultimately, how is FLVCR2-mediated heme transport linked to cerebral angiogenesis? To answer these questions, Dr. Cater has formed a comprehensive research plan combining structural biology, biophysics, membrane protein biochemistry, electrophysiology, and cellular biology. It involves cryo-EM structure determination of FLVCR2; functional characterization of FLVCR2 using liposome-based assays and electrophysiology; interrogation of the link between FLVCR2 and angiogenesis using brain endothelial cell-based assays; and mapping the FLVCR2 interactome using proximity-based labeling assays. This research will provide key insights into how FLVCR2 transports heme in brain endothelial cells and this in turn regulates angiogenesis. The proposed studies for the K99 phase will largely take place at Columbia University, which is home to a vibrant and collegial community of structural biologists, biophysicists, and biochemists. This environment is ideal to facilitate Dr. Cater in the successful completion of the proposed K99 research program and achieving her goal of transitioning into a successful independent researcher.

项目摘要 此应用程序详细介绍了职业发展和研究计划，这些计划是专门为促进 首席研究员迷迭香J.卡特博士转变为独立的学术职位。卡特医生 具有多学科背景，并通过这个K99/R 00提案，寻求最终确定她的生化 培训，掌握一个新系统的生物学，并为她作为一名独立调查员的职业生涯获得技能。 该奖项的K99阶段（前两年）涉及一个结构化的职业发展计划，将允许博士。 迎合：提高她在单粒子冷冻电子显微镜（cryo-EM）方面的技能;获得大脑方面的基本培训 基于内皮细胞的测定和基于邻近的标记实验;并获得重要的职业技能 例如，撰写拨款、科学交流、实验室管理、教学、负责任的行为， 研究，并掌握学术工作申请流程。职业发展计划还包括明确的 以及确定并成功获得独立终身教职的可行步骤， K99阶段的结束。卡特博士组建了一支由多学科导师、顾问和 合作者，将监督和指导她的培训，研究计划，并过渡到独立。 建议的研究计划涵盖了该奖项的指导（K99）和独立（R 00）阶段。它 涉及血红素转运蛋白FLVCR 2的机制研究，FLVCR 2在血脑屏障内表达 内皮细胞，并在脑血管系统的发育中起关键作用。K99的研究计划 阶段旨在在分子水平上表征FLVCR 2的结构和功能。R 00研究计划 然后旨在描述FLVCR 2如何在细胞水平上调节血管生成。核心研究问题 解决包括：什么是重要的分子决定因素FLVCR 2结合血红素特异性？什么 FLVCR 2转运血红素必须进行构象变化吗？是什么推动了这一运输过程？和 最终，FLVCR 2介导的血红素转运如何与脑血管生成相关？ 为了回答这些问题，卡特博士形成了一个综合性的研究计划， 生物物理学、膜蛋白生物化学、电生理学和细胞生物学。它涉及到冷冻电镜结构 FLVCR 2的测定;使用基于脂质体的测定法对FLVCR 2进行功能表征， 电生理学;使用脑内皮细胞为基础的FLVCR 2和血管生成之间的联系的询问 使用基于邻近性的标记测定对FLVCR 2相互作用组作图。这项研究将提供 FLVCR 2如何在脑内皮细胞中转运血红素，进而调节血管生成的关键见解。 K99阶段的拟议研究将主要在哥伦比亚大学进行，该大学拥有一个充满活力的 以及由结构生物学家、生物化学家和生物化学家组成的学院团体。这种环境非常适合 协助Cater博士成功完成拟议的K99研究计划并实现其目标 成为一名成功的独立研究者