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

• 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
• 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

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

项目摘要 该建议是针对乔恩·威斯勒（Jon Wisler）博士的五年研究计划， 创伤，重症监护和烧伤手术。该建议旨在研究外泌体介导的影响 在约翰·克里斯曼（John Christman）博士的心态下，在败血症期间发生的表观遗传调节。克里斯曼博士是 单核细胞/巨噬细胞生物学和表观遗传学领域的高产研究人员 心态和生产力的迹象。提案的研究和职业发展部分着重于 技术扩展和专业发展。这包括一个精致的逻辑计划，可测量的短 和长期里程碑。利用他的Mentalship团队的丰富经验，该建议将培训Jon 是尖端技术，包括CRISPR-GENE编辑，活细胞成像，纳米颗粒RNA包装和 分娩和外泌体生物学可大大提高他的机械理解和研究能力。 败血症患者表现出深度的免疫抑制，随后的传染性较高 并发症和长期死亡率增加。我们的初步数据确定了DNA的显着增加 脓毒症患者循环外泌体中的甲基转移酶（DNMT）mRNA，该DNMT mRNA 从这些外泌体转移到幼稚的单核细胞。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。我们对此应用的目的是阐明 外部摄取，并在体内败血症模型中靶向这些表观遗传事件。我们假设预防 外泌体介导的DNA甲基化可以在败血症过程中维持宿主免疫反应。 总体目的是确定基于外部介导的表观遗传控制的机械基础 控制败血症相关免疫抑制的事件，并确定靶向的潜在翻译价值 DNMT功能以治疗与败血症相关的并发症。结合新颖，尖端的培训 技术将大大提高Jon的科学专业知识，并允许从Mendored到 独立的外科医生科学家。 目标1：阐明外泌体细胞摄取的机制并定义了 DNMT作为表观遗传信使，有助于后塞介导的免疫抑制 目标2：确定靶向外泌体介导的抗DNMT递送的安全性和效率 小鼠模型中败血症期间的免疫抑制