Mechanisms of Endothelial Cell Dysfunction in Critically Ill Children

危重儿童内皮细胞功能障碍的机制

基本信息

批准号：
10450069
负责人：
RICHARD W PIERCE
金额：
$ 13.81万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10450069
关键词：
Adherens Junction Bioinformatics Biological Assay Biological Models Biology Blood Blood Vessels Blood capillaries CRISPR/Cas technology Candidate Disease Gene Capillary Endothelial Cell Capillary Leak Syndrome Capillary Permeability Cardiopulmonary Cell Culture Techniques Cell Line Cells Cessation of life Child Childhood Clinical Critical Care Critical Illness Critically ill children Data Dermal Deterioration Development Edema Electrical Resistance Endothelial Cells Endothelium Etiology Event Extramural Activities Failure Fellowship Functional disorder Funding GTPase-Activating Proteins Gene Expression Gene Mutation Genes Genetic Genetic Techniques Genetic Transcription Goals Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Human Immunologic Techniques Immunology Individual Inflammatory Jordan Laboratories Liquid substance Lung Mediating Mediator of activation protein Medicine Mentors Mentorship Methodology Methods Modeling Modification Monomeric GTP-Binding Proteins Morbidity - disease rate Multiple Organ Failure Mutation Nutrient Organ Pathologic Pathway interactions Patients Permeability Pharmacology Play Process Protocols documentation Regulation Research Research Methodology Research Personnel Research Project Grants Research Support Resolution Resources Role Scientist Signal Pathway Signal Transduction Skin Syndrome TNF gene Testing Tight Junctions Time Training Umbilical vein Vascular Permeabilities Venous Work cell type clinically significant cytokine endothelial dysfunction experience gene product improved instructor interstitial knock-down lung microvascular endothelial cells lymphotoxin beta medical schools mortality multidisciplinary novel novel therapeutic intervention overexpression pediatric patients prevent professor programs response single-cell RNA sequencing skills symposium targeted treatment therapy development tool transcriptome

项目摘要

Project Summary Candidate. I am an Instructor in pediatric critical care medicine at Yale School of Medicine and will be promoted to Assistant Professor with academic promotion per Yale School of Medicine's Clinician Scholar track in July 2018. My K08 application will allow me to acquire additional mentored research experience so that I may become an independent investigator with expertise in endothelial cell signaling and its disruption in critically ill children. This research will build on my fellowship training that focused on the signals that regulate changes in vascular permeability and their associated clinical consequences and will further develop my laboratory skills required to successfully conduct vascular research, such as genetic modification of human cells, interrogation of intracellular signaling pathways, and bioinformatic analysis. I will take graduate courses on vascular biology, bioinformatics, and immunology research methods and will participate in national conferences. I have an outstanding mentor, Dr. Jordan Pober, a world-class vascular biologist, along with advisors Dr. Mustafa Khokha, expert in genetics, and Dr. Vince Faustino, expert biostatistician and clinical trialist. This multidisciplinary mentorship team, along with the resources available through the VBT program at Yale, will allow me to develop the necessary skills to function as an extramurally funded clinician scientist focused on improving the care of critically ill children. Research Project. Endothelial cells (ECs) actively regulate vessel permeability that is essential for organ function and patient survival. During cardiopulmonary failure, loss of EC permselectivity results in capillary leak that contributes to morbidity and mortality in children. The unique regulation of the signaling events that produce permeability changes in capillary EC is largely unknown. Likewise, how critical illness induces EC dysfunction remains unknown, due in large part to our inability to directly assess EC changes in critically ill patients. I recently demonstrated the importance of the regulation of the small GTPase RhoB in a pediatric patient with systemic capillary leak syndrome caused by a single gene mutation. In Aim 1, I test the hypothesis that pathological capillary permeability is regulated by small GTPases and that these pathways are influenced by transcriptional changes induced by inflammatory cytokines. I will interrogate intracellular signaling pathways in capillary ECs cultured from different organs (skin and lung) to determine the effect of their modulation on trans-endothelial electrical resistance. In Aim 2, I will test the hypothesis that ECs undergo transcriptional changes in response to critical illness. I will analyze the entire transcriptome of individual venous ECs collected directly from critically ill children using single cell RNA sequencing. I will assess the functions of identified candidate gene products using pharmacologic, genetic, and immunologic techniques to manipulate and analyze their effect on the intracellular signaling pathways that control the initiation and resolution of capillary leak. Better understanding the pathways involved in the development and resolution of capillary leak is the first step in developing therapies for this common condition associated with serious morbidity and mortality.

项目摘要候选人。我是耶鲁大学医学学院儿科重症监护医学的讲师，并将晋升七月，耶鲁大学医学学院的临床医生曲目学术晋升助理教授 2018年。我的K08应用程序将使我能够获得更多的指导研究经验，以便我成为具有内皮细胞信号传导专业知识及其在重病儿童中的破坏方面具有专业知识的独立研究者。这项研究将基于我的奖学金培训，该培训的重点是调节血管变化的信号渗透性及其相关的临床后果，并将进一步发展我所需的实验室技能成功进行血管研究，例如人类细胞的遗传修饰，细胞内询问信号通路和生物信息学分析。我将参加有关血管生物学，生物信息学的研究生课程，和免疫学研究方法，并将参加民族会议。我有一位出色的导师，世界一流的血管生物学家乔丹·珀伯（Jordan Pober）博士以及顾问Mustafa Khokha博士，遗传学专家，以及文斯·福斯蒂诺（Vince Faustino）博士，专家生物统计学家和临床试验师。这个多学科的指导团队，通过耶鲁大学的VBT计划获得的资源，我将允许我发展必要的技能作为一名临床医生的临床医生，专注于改善重症儿童的护理。研究项目。内皮细胞（EC）主动调节器官必不可少的血管通透性功能和患者生存。在心肺衰竭期间，EC允许选择性的丧失导致毛细管泄漏这有助于儿童的发病率和死亡率。产生的信号事件的独特调节毛细管EC的渗透性变化在很大程度上未知。同样，重症疾病如何引起EC功能障碍仍然未知，这在很大程度上是由于我们无法直接评估重症患者的EC变化。我最近证明了针对全身性的小儿患者调节小型GTPase RHOB的重要性由单个基因突变引起的毛细血管泄漏综合征。在AIM 1中，我检验了病理学的假设毛细管渗透性受小的GTPases调节，这些途径受转录的影响炎症细胞因子引起的变化。我将询问毛细管EC中的细胞内信号通路从不同的器官（皮肤和肺）培养以确定其调节对跨内皮的影响电阻。在AIM 2中，我将检验以下假设：EC经历转录的变化。重症病。我将直接分析直接从重病收集的单个静脉EC的整个转录组使用单细胞RNA测序的儿童。我将使用药理，遗传和免疫学技术来操纵和分析其对细胞内的影响控制毛细管泄漏的启动和分辨率的信号通路。更好地了解路径参与毛细管泄漏的开发和解决是为此开发疗法的第一步与严重发病率和死亡率相关的常见状况。