Inflammation and Cerebral Aneurysm Formation: Targets for Modulation and Vascular

项目总结/摘要 候选人：我的长期职业目标是成为脑动脉瘤研究领域的领先临床科学家。我在斯坦福大学读本科时，在Edward Mocarksi博士的实验室工作，研究人类巨细胞病毒B2.7启动子的调控，从那时起，我就长期致力于生物医学研究。作为哥伦比亚大学的一名医学生，我在扫罗西尔弗斯坦博士的指导下绘制了单纯疱疹病毒IE 0基因的功能域。我在E博士的指导下工作。Sander Connolly和大卫平斯基博士在小鼠模型中研究中风的病理生理机制。作为 我是马萨诸塞州总医院的住院医生和研究员，在约瑟夫·瓦坎蒂博士的实验室工作，研究一个用内皮细胞接种聚合物的项目，以创造一种治疗脑动脉瘤的血管移植物。我未来五年的目标是成为一名独立调查员。我将开始一个严格的教学计划，重点是获得必要的知识和技能，以实现这一目标。我将在获奖期间获得的科学技能，包括动物外科，组织学，免疫组织化学，活体显微镜，干细胞生物学技术和先进的数据统计分析。为了补充我的实验室工作经验，我将在年参加佛罗里达大学跨学科项目的研究生课程。 生物医学科学我已经制定了一个全面的职业发展计划，其中包括实验室培训，密切指导，研究生水平的课程，研究会议和口头报告。我的目标是在颁奖结束时获得NIH R 01奖。 工作环境：我将在爱德华·斯科特博士的实验室工作，他是一位杰出的导师，曾成功培训过几位临床科学家和K 08奖的获奖者。我会请材料科学与工程系的克里斯托弗·巴蒂奇博士作为顾问，他是一位出色的合作者。我将在一个优秀的科学和创造性的环境中与干细胞生物学和再生医学，麦克奈特脑研究所和材料科学与工程系的合作者一起工作，以指导我的研究进展。我得到了我的主席、神经外科系和佛罗里达大学的全力支持，我将至少50%的时间和精力投入到我的研究职业发展中。所有的组成部分都到位，以促进我的研究建议和我的整体研究职业发展的成功结果。 研究计划：该项目的长期目标是更好地定义脑动脉瘤形成的病理生理学，从而开发治疗脑动脉瘤的转化疗法。总体目标是回答炎症是否导致脑动脉瘤形成的知识的根本差距，或者是一种附带现象;识别可能是动脉瘤炎症反应调节靶点的信号传导机制;并识别可以修复血管炎症退行性机制的血管修复细胞。中心假设是：脑动脉瘤形成是一个炎症过程。调节炎症反应将改变脑动脉瘤的发展。可以识别出血管修复细胞，它们可以修复血管炎症引起的退行性机制。这项研究的基本原理是，由于脑动脉瘤可以在破裂前被诊断出来，因此修复动脉瘤炎症过程并防止动脉瘤进展到破裂的治疗将对公众健康产生重大有益影响。在强有力的初步数据的指导下，将通过追求三个具体目标来测试该假设：1）证明脑动脉瘤形成是由炎症过程引起的; 2）识别炎症过程中涉及的信号传导机制，作为调节的靶点，以改变动脉瘤的发展; 3）识别修复血管炎症引起的退行性机制的血管修复细胞。在第一个目标下，将进行两条线的实验：1）通过激活炎症（将活化的巨噬细胞和嗜中性粒细胞注射到动脉瘤中）在小鼠中产生动脉瘤。 正常小鼠颈动脉）;和2）在不能产生炎症反应的免疫缺陷小鼠中进行弹性蛋白酶囊状动脉瘤模型。在第二个目标下，白细胞募集级联每个步骤的信号机制：1）趋化性，2）滚动，3）粘附，4）迁移;将在弹性蛋白酶囊性动脉瘤模型中被阻断。在第三个目标下，CD 133+内皮祖细胞（EPC），造血干细胞和间充质干细胞的细胞特异性群体将被接种到Matrigel和我们的合作者Christopher Batich博士开发的其他聚合物中，并注射到小鼠弹性蛋白酶囊状动脉瘤中，随后将评估再内皮化和抑制血管炎症反应的证据。

PROJECT SUMMARY/ABSTRACT Candidate: My long-term career goal is to become a leading clinician-scientist in the field of cerebral aneurysm research. I have had a long commitment to biomedical research since an undergraduate at Stanford University working in the laboratory of Dr. Edward Mocarksi on a project studying regulation of the B2.7 promoter of human cytomegalovirus. As a medical student at Columbia University I worked under Dr. Saul Silverstein mapping functional domains of the IE0 gene of herpes simplex virus. I then worked under Dr. E. Sander Connolly and Dr. David Pinsky studying pathophysiologic mechanisms of stroke in a mouse model. As a resident and fellow at the Massachusetts General Hospital, I worked in the laboratory of Dr. Joseph Vacanti on a project seeding polymers with endothelial cells to create a vascular graft for treating cerebral aneurysms. My immediate objective for the next five years is to become an independent investigator. I will start a rigorous didactic program focused on gaining the knowledge and skills necessary to achieve this objective. Scientific skills I will acquire during the award period, include animal surgery, histology, immunohistochemistry, intravital microscopy, stem cell biology techniques, and advanced statistical analysis of data. To complement my laboratory experience, I will take graduate coursework in the University of Florida Interdisciplinary Program in Biomedical Sciences. I have developed a comprehensive career development program which includes laboratory training, close mentorship, graduate level coursework, research conferences, and oral presentations. My goal is to obtain an NIH R01 award by the time the award finishes. Environment: I will be working in the laboratory of Dr. Edward Scott, an outstanding mentor, who has successfully trained several clinician-scientists and previous recipients of the K08 award. I will have as a consultant, Dr. Christopher Batich in the Department of Materials Science and Engineering, a superb collaborator. I will be working in an outstanding scientific and creative environment with collaborators in the Program in Stem Cell Biology and Regenerative Medicine, the McKnight Brain Institute, and the Department of Materials Science and Engineering to guide my research progress. I have the full committed support of my chairman, the Department of Neurological Surgery, and the University of Florida to devote at least 50% of my time and effort to my research career development. All of the components are in-place to promote a successful outcome in my research proposal and my overall research career development. Research Plan: The long-term goal of this project is to better define the pathophysiology of cerebral aneurysm formation, and, thereby, develop a translational therapy that would treat cerebral aneurysms. The overall objective is to answer the fundamental gap in knowledge of whether inflammation causes cerebral aneurysm formation, or is an epiphenomenon; identifying signaling mechanisms which could be targets for modulation of the aneurysm inflammatory response; and identifying vascular repair cells that can repair the degenerative mechanisms of vascular inflammation. The central hypothesis is that: Cerebral aneurysm formation is an inflammatory process. Modulating the inflammatory response would alter the development of cerebral aneurysms. Vascular repair cells can be identified that repair the degenerative mechanisms caused by vascular inflammation. The rationale for the proposed research is that because cerebral aneurysms can be diagnosed before they rupture, a therapy that would repair the aneurysm inflammatory process and prevent aneurysms from progressing to rupture, would have a significant beneficial impact for the public health. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: 1) Demonstrate that cerebral aneurysm formation is caused by an inflammatory process; 2) Identify signaling mechanisms involved in the inflammatory process as targets for modulation to alter the development of aneurysms; and 3) Identify vascular repair cells that repair the degenerative mechanisms caused by vascular inflammation. Under the first aim, two lines of experiments will be performed: 1) aneurysms will be produced in mice by activating inflammation (injecting activated macrophages and neutrophils into normal mouse carotid artery); and 2) performing an elastase saccular aneurysm model in immune-deficient mice that can not mount an inflammatory response. Under the second aim, signaling mechanisms at each step of the leukocyte recruitment cascade: 1) chemotaxis, 2) rolling, 3) adhesion, 4) transmigration; will be blocked in an elastase saccular aneurysm model. Under the third aim, cell-specific populations of CD133+ endothelial progenitor cells (EPCs), hematopoietic stem cells, and mesenchymal stem cells will be seeded into Matrigel and other polymers developed by our collaborator, Dr. Christopher Batich, and injected into murine elastase saccular aneurysms which will later be evaluated for evidence of re-endothelialization and inhibition of the vascular inflammatory response.