UTILIZING SOLUBLE VIMENTIN AND ITS COMPONENTS TO ATTENUATE INFLAMMATION

利用可溶性波形蛋白及其成分来减轻炎症

基本信息

批准号：
9889964
负责人：
FONG WILSON LAM
金额：
$ 18.02万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9889964
关键词：
Acute Lung Injury Address Adhesions Adult Respiratory Distress Syndrome Advisory Committees Animals Attenuated Award Binding Biochemistry Biological Assay Biology Biometry Blood Platelets Cell Adhesion Molecules Cell surface Cells Childhood Circular Dichroism Clinical Complex Critical Care Critical Illness Cytoskeleton Data Development Devices Distant Endothelial Cells Endothelium Endotoxemia Enrollment Experimental Designs Experimental Models Functional disorder Funding Future Goals Health Human In Vitro Incidence Infection Inflammation Inflammation Process Injury Intermediate Filament Proteins Intracellular Transport Kinetics Knowledge Laboratories Lead Leukocyte Rolling Leukocyte Trafficking Leukocytes Life Lung Lung Inflammation Maintenance Measures Mediating Medicine Mentors Mesenchymal Microcirculation Modeling Molecular Biology Morbidity - disease rate Mus Organ Outcome P-Selectin P-selectin ligand protein Pathologic Pathology Patients Pediatrics Persons Pharmacology Physical activity Physicians Plasma Play Postdoctoral Fellow Protein Biochemistry Publications Publishing Recombinant Proteins Recombinants Reperfusion Injury Reporting Research Research Ethics Research Personnel Role Scientist Sepsis Site Structure Surface Surface Plasmon Resonance Technical Expertise Testing Therapeutic Thrombosis Time Training Programs United States United States National Institutes of Health Vascular Endothelium Vimentin Writing biophysical techniques career cecal ligation puncture college improved improved outcome interest intravital microscopy lung injury migration mortality mouse model neutrophil novel novel therapeutics pre-clinical prevent professor receptor recruit safety study tissue injury venule wound healing

项目摘要

Project Abstract I am an Assistant Professor of Pediatrics at the Baylor College of Medicine (BCM) with a long- standing interest in the complex interaction between neutrophils, platelets, and endothelium in the microcirculation. As a Pediatric Critical Care Medicine physician, my main career goal is to become a successful independent investigator with a focus on pathology inflammation and how it contributes to the morbidity and mortality in critically-ill patients, such as in acute lung injury and sepsis (severe infection). As a post-doctoral clinical fellow, I investigated the proinflammatory effect of platelets on neutrophil transmigration that lay the groundwork for this proposal. The primary objective of this K08 proposal is to obtain additional expertise in protein biochemistry and animal studies to be able to discover and test novel therapies for these devastating conditions. This proposal will allow me to have protected time to attend classes in molecular biology and biochemistry as well as hone my technical skills in the laboratory. Additionally, it will allow me to enroll in the Clinical Scientist Training Program at BCM, which will improve my knowledge in experimental design, scientific writing, research ethics, and biostatistics. My primary research mentor, Dr. Miguel Cruz, is a renowned researcher with successful funding on studying microvascular thrombosis in systemic inflammation. He has been successful in creating recombinant proteins, including the soluble vimentin used in this proposal. In addition to Dr. Cruz, my research advisory committee consists of Drs. Timothy Palkzill (Pharmacology), David Corry (Murine models of lung inflammation), C Wayne Smith (leukocyte trafficking), and Perumal Thiagarajn (platelet biology), who are all experts in their respective fields related to this proposal. We have observed that soluble vimentin decreases inflammation by blocking leukocyte adhesion to both platelets and endothelial cells through blocking the interaction between P-selectin and its counter-part, P-selectin glycoprotein ligand-1 (PSGL-1). Our preliminary studies in mice suggest that soluble vimentin decreases injury to the lung in one model of sepsis. Whether soluble vimentin results in decreasing lung injury and mortality due to sepsis is unknown. My proposal will test the central hypothesis that soluble vimentin attenuates inflammation by blocking P-selectin-PSGL-1 interactions to decrease leukocyte adhesion and transmigration across endothelium. I will test this hypothesis through the following 3 specific aims: (1) Identify the active motif and binding kinetics of vimentin-P- selectin interactions, (2) Evaluate the effect of soluble vimentin on platelet-enhanced neutrophil transmigration across inflamed endothelium in vitro, and (3) Evaluate the efficacy of soluble vimentin, and its components, in reducing secondary acute lung injury in mice in two models of experimental sepsis, endotoxemia and cecal-ligation and perforation. In Aim 1, I will use different biophysical techniques, such as surface plasmon resonance and circular dichroism, to evaluate the structure and binding kinetics of soluble vimentin to P-selectin. In Aim 2, I will use in vitro dynamic and static adhesion assays to measure the effect of soluble vimentin on neutrophil transendothelial migration, a novel observation on which I had previously published. Finally, in Aim 3, I will study two different models of experimental sepsis because they have different mechanisms of inflammation. I will also study the safety of soluble vimentin administration. I anticipate that soluble vimentin will attenuate inflammation by blocking leukocyte-platelet-endothelial interactions and that it will decrease lung injury in sepsis and improve mortality, which will lead to novel therapies to prevent and treat critically ill patients.

项目摘要我是贝勒医学院（BCM）的儿科助理教授对中性粒细胞，血小板和内皮之间的复杂相互作用的兴趣微循环。作为一名儿科重症监护医学医师，我的主要职业目标是成为一名成功的独立研究者，专注于病理炎症以及如何它有助于急诊患者的发病率和死亡率，例如急性肺损伤和败血症（严重感染）。作为博士后临床研究员，我调查了血小板对中性粒细胞传播的促炎作用，为此奠定了基础提议。该K08提案的主要目的是获得蛋白质方面的额外专业知识生物化学和动物研究能够发现和测试这些新疗法毁灭性的条件。该建议将使我有受保护的时间参加上课分子生物学和生物化学以及在实验室中磨练我的技术技能。此外，这将使我能够参加BCM的临床科学家培训计划，该计划将提高我在实验设计，科学写作，研究伦理和生物统计学。我的主要研究导师Miguel Cruz博士是一位著名的研究员在研究系统性炎症中研究微血管血栓形成方面的成功资金。他有成功地创建了重组蛋白，包括此中使用的可溶性波形蛋白提议。除克鲁兹博士外，我的研究咨询委员会还包括博士。蒂莫西 Palkzill（药理学），David Corry（肺部炎症的鼠模型），C Wayne Smith （白细胞贩运）和Perumal Thiagarajn（血小板生物学），他们都是专家与此提案有关的各个领域。我们观察到可溶性波形蛋白会降低通过阻断白细胞粘附到血小板和内皮细胞的炎症阻止P-选择素与其反处理之间的相互作用P-选择蛋白糖蛋白配体-1 （PSGL-1）。我们在小鼠中的初步研究表明，可溶性波形蛋白减少了对肺中的肺部模型。可溶性波形蛋白是否会减少肺损伤和由于败血症而导致的死亡率尚不清楚。我的建议将检验可溶性的中心假设波形蛋白通过阻止p-选择蛋白-PSGL-1相互作用来减少炎症白细胞粘附和跨内皮的移民。我将通过以下3个特定目的：（1）确定波形蛋白-p-的主动基序和结合动力学选择素相互作用，（2）评估可溶性波形蛋白对血小板增生的影响体外跨越内皮的迁移，（3）评估可溶性的功效波形蛋白及其成分在减少两种模型中小鼠的继发性急性肺损伤中实验性败血症，内毒素血症以及盲肠结合和穿孔。在AIM 1中，我会使用不同的生物物理技术，例如表面等离子体共振和圆形二色性，评估可溶性波形蛋白与P-选择素的结构和结合动力学。在AIM 2中，我会使用体外动态和静态粘附测定，以测量可溶性波形蛋白对中性粒细胞跨内皮迁移，这是我以前发表的新颖观察。最后，在AIM 3中，我将研究两种不同的实验败血症模型，因为它们具有炎症的不同机制。我还将研究可溶性波形蛋白的安全性行政。我预计可溶性波形蛋白会通过阻塞来减轻炎症白细胞 - 平台 - 内皮相互作用，它将减少败血症的肺损伤提高死亡率，这将导致新的疗法预防和治疗重症患者。