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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要该申请详细介绍了专门定制的职业发展和研究计划，以促进首席研究员 Rosemary J. Cater 博士转任独立学术职务。卡特博士具有多学科背景，并通过此 K99/R00 提案，寻求最终确定她的生化培训，掌握新系统的生物学，并获得作为独立调查员的职业技能。该奖项的 K99 阶段（前两年）涉及一个结构化的职业发展计划，该计划将使 Dr. 致力于：提高她在单粒子冷冻电子显微镜（cryo-EM）方面的技能；获得必要的大脑训练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拨款写作、科学传播、实验室管理、教学、负责任的行为研究和掌握学术工作申请流程。职业发展规划还包括明确的以及确定并成功获得独立终身教授职位的可行步骤 K99阶段结束。 Cater 博士组建了一支由多学科导师、顾问和专家组成的顶级团队合作者将监督和指导她的培训、研究计划以及向独立的过渡。提出的研究计划涵盖该奖项的指导阶段（K99）和独立阶段（R00）。它涉及血红素转运蛋白 FLVCR2 的机制研究，该转运蛋白在血脑屏障内表达内皮细胞在脑血管系统的发育中起着关键作用。 K99的研究计划阶段的目的是在分子水平上表征FLVCR2的结构和功能。 R00研究计划然后旨在描述 FLVCR2 如何在细胞水平上调节血管生成。核心研究问题解决的问题包括：FLVCR2 特异性结合血红素的重要分子决定因素是什么？什么 FLVCR2 必须进行构象变化才能运输血红素吗？是什么驱动了这个运输过程？和最终，FLVCR2 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，Cater博士结合结构生物学、生物物理学、膜蛋白生物化学、电生理学和细胞生物学。它涉及冷冻电镜结构 FLVCR2的测定；使用基于脂质体的测定法对 FLVCR2 进行功能表征电生理学；使用基于脑内皮细胞的方法询问 FLVCR2 与血管生成之间的联系化验；并使用基于邻近的标记分析绘制 FLVCR2 相互作用组图。这项研究将提供关于 FLVCR2 如何在脑内皮细胞中转运血红素并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，该大学是充满活力的研究中心的所在地以及结构生物学家、生物物理学家和生物化学家的学院社区。这种环境非常适合协助 Cater 博士成功完成拟议的 K99 研究计划并实现她的目标转变为一名成功的独立研究员。