Analyzing the mechanism of exosome mediated DNA Methyltransferase activity during sepsis

分析脓毒症期间外泌体介导的 DNA 甲基转移酶活性机制

基本信息

批准号：
10591480
负责人：
Jon R Wisler
金额：
$ 9.73万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10591480
关键词：
Ablation Affect Animal Model Area Biochemical Biology Cell Culture Techniques Cell physiology Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coupled Critical Care DNA Methylation DNA Modification Methylases DNA delivery Data Defense Mechanisms Development Development Plans Disease Epigenetic Process Event Exhibits Exposure to Gene Silencing Genes Goals Human Immune Immune System Diseases Immune response Immunology Immunosuppression In Vitro Incubated Infection Inflammatory Interleukin-1 beta Interleukin-6 Interruption Intervention Label Macrophage Maintenance Measurable Mediating Membrane Fusion Mentors Mentorship Messenger RNA Methylation Molecular Mus Nanotechnology Operative Surgical Procedures Outcome Pathway interactions Patients Persons Phagocytes Phagocytosis Phenotype Predisposition Prevention Principal Investigator Production Productivity Protein Isoforms RNA Regulation Research Research Personnel Role Safety Scientist Sepsis Signal Transduction Small Interfering RNA Surgeon Survivors TNF gene Techniques Technology Testing Training Trauma base career career development combat cytokine efficacy evaluation epigenetic regulation exosome experience genetic manipulation immune function improved in vivo in vivo Model insight interdisciplinary approach knock-down live cell imaging monocyte mortality mouse model nanoparticle novel novel therapeutics pharmacologic prevent professor programs promoter research and development response septic siRNA delivery trafficking translational potential trauma care uptake wortmannin

项目摘要

Project Summary This proposal is for a five-year research program for Dr. Jon Wisler, an Assistant Professor in the Division of Trauma, Critical Care, and Burn Surgery. This proposal aims to study the effects of exosome-mediated epigenetic regulation that occurs during sepsis, under the mentorship of Dr. John Christman. Dr. Christman is a highly productive researcher in the fields of monocyte/macrophage biology and epigenetics who has a long track of mentorship and productivity. The research and career development portions of the proposal focus on technique expansion and professional development. This includes a refined, logical plan with measurable short and long-term milestones. Utilizing the extensive experience of his mentorship team, this proposal will train Jon is cutting-edge technologies including CRISPR-gene editing, live-cell imaging, nanoparticle RNA packaging and delivery, and exosome biology to greatly improve his mechanistic understanding and investigatory capabilities. Patients with sepsis exhibit a profound degree of immunosuppression with higher levels of subsequent infectious complications and increased long-term mortality. Our preliminary data identifies significant increases in DNA Methyltransferases (DNMT) mRNAs in circulating exosomes of patients with sepsis, and that this DNMT mRNA is transferred from these exosomes to naïve monocytes. This transfer results in increased epigenetic events (promoter methylation) and gene silencing. Our intent for this application is to elucidate the mechanisms of exosome uptake, and target these epigenetic events in an in vivo model of sepsis. We hypothesize prevention of exosome-mediated DNA methylation allows for maintenance of the host immune response during sepsis. The overall objective is to identify the mechanistic base that underlies exosome-mediated control of the epigenetic events that govern sepsis-related immunosuppression, and identify the potential translational value of targeting DNMT function to treat complications associated with sepsis. Incorporating the training of novel, cutting-edge techniques will greatly improve the Jon's scientific expertise, and allow for the progression from mentored to independent surgeon-scientist. Aim 1: Elucidate the mechanisms that underlie the cellular uptake of exosomes and define the role of DNMT as an epigenetic messenger that contributes to post-sepsis mediated immunosuppression Aim 2: Establish the safety and efficacy of targeting exosome-mediated delivery of anti-DNMT to prevent immunosuppression during sepsis in mouse models

项目概要该提案是为乔恩·威斯勒博士（Jon Wisler）进行的为期五年的研究计划，他是该部门的助理教授。该提案旨在研究外泌体介导的外伤、重症监护和烧伤手术的影响。在约翰·克里斯特曼博士的指导下，败血症期间发生的表观遗传调控是。单核细胞/巨噬细胞生物学和表观遗传学领域高产的研究人员，拥有长期的研究经验提案的研究和职业发展部分重点关注指导和生产力。这包括一个精致的、有逻辑的、可衡量的短期计划。该提案将利用他的指导团队的丰富经验来培训乔恩。是尖端技术，包括 CRISPR 基因编辑、活细胞成像、纳米粒子 RNA 包装和递送和外泌体生物学，极大地提高了他的机制理解和研究能力。脓毒症患者表现出严重的免疫抑制，随后的感染水平较高我们的初步数据表明 DNA 显着增加。脓毒症患者循环外泌体中的甲基转移酶 (DNMT) mRNA，并且该 DNMT mRNA 从这些外泌体转移到幼稚单核细胞，这种转移导致表观遗传事件增加。（启动子甲基化）和基因沉默我们的目的是阐明其机制。外泌体的摄取，并在脓毒症体内模型中针对这些表观遗传事件，我们勇敢地进行了预防。外泌体介导的 DNA 甲基化可以在脓毒症期间维持宿主免疫反应。总体目标是确定外泌体介导的表观遗传控制的机制基础控制脓毒症相关免疫抑制的事件，并确定靶向的潜在转化价值 DNMT 功能可治疗脓毒症相关并发症。技术将极大地提高乔恩的科学专业知识，并允许从指导到发展独立外科医生兼科学家。目标 1：阐明细胞摄取外泌体的机制并定义外泌体的作用 DNMT 作为表观遗传信使，有助于脓毒症后介导的免疫抑制目标 2：确定外泌体介导的抗 DNMT 靶向递送的安全性和有效性，以预防小鼠模型败血症期间的免疫抑制