Altered Treg Differentiation in ALD: A Novel Role for Anaphylatoxins C3a and C5a

项目总结 该候选人是一名博士后研究员和致力于发展学术事业的年轻研究员。 重点研究了乙醇诱导的分子机制的鉴定和表征 炎症，导致肝脏耐受性受损和组织损伤。有很强的氧化背景 应激和先天免疫与酒精性肝损伤有关，候选人已经形成了特别的 在使用老鼠模型进行拟议研究方面的专业知识。这些模型系统的使用有 补体和过敏性毒素在介导肝脏炎症和损伤中的重要作用 由乙醇引起。此外，我们假设Treg动态平衡改变是受损的致病事件。 消退炎症和损伤。候选人最近和目前的工作为她提供了 她有机会发展自己的研究项目，并开始向独立过渡。职业生涯 建议书中描述的发展计划概述了为期2年的指导性培训，包括技术培训 除了职业发展活动之外的技能培训，旨在促进成功过渡到 独立。成功后的为期3年的自主科学和职业发展计划 还概述了助理教授职位的招聘情况。候选人的导师有一条经过验证的轨迹- 优秀的科学生产力和成功的指导记录，可以为应聘者提供坚实的 她在克利夫兰诊所勒纳研究所的实验室里的研究环境。研究计划： 酒精性肝病(ALD)仍然是主要的社会经济负担。几十年来，发病率和 死亡率保持不变，患者的长期预后仍然很差。ALD是多因素的， 部分是由先天免疫系统调节的。补体，先天免疫的一个组成部分，也是 与肌萎缩侧索硬化症的发生发展有关，并正在成为一个有吸引力的治疗靶点。我们早期的研究已经 发现异常的、不受控制的补体激活会导致组织损伤；然而，补体 也是肝脏愈合所必需的。这些研究为我们目前的提案提供了基础：特定于细胞 补体疗法是目前抑制疗法的必要替代疗法，可以作为一种新的治疗方法 ALD的治疗。调节性T细胞(Treg)是炎症和炎症分解阶段的关键贡献者 通过分泌大量免疫抑制和促进分解的细胞因子来维持免疫耐受性。 晚期ALD患者外周血Treg减少，但乙醇改变T细胞的机制 动态平衡尚未得到研究。我们的工作假设是持续的补体激活 酒精导致肝脏中Treg的异常调节，从而导致肝脏炎症和组织 在三个特定的目标中，我们将表征乙醇介导的Treg在以下肝脏中的分化 小鼠高脂饮食，探讨过继转移体外分化iTreg的能力 减轻酒精喂养小鼠的肝脏炎症和减轻肝损伤，并评估细胞外小泡对 刺激幼稚的CD4+T细胞体外分化为iTreg和Th17亚型。我们期待的结果是 这些目标将为未来乙醇的机理研究和临床干预提供坚实的基础。 引起炎症和组织损伤。

PROJECT SUMMARY The Candidate is a postdoctoral fellow and young investigator dedicated to developing an academic career focused on the identification and characterization of molecular mechanisms stemming from ethanol-induced inflammation, contributing to impaired liver tolerance and tissue injury. With a strong background in oxidative stress and innate immunity related to ethanol-induced liver injury, the candidate has developed particular expertise in the use of mouse models to conduct the proposed studies. Use of these model systems has revealed and important role for complement, and anaphylatoxins in mediating liver inflammation and injury caused by ethanol. Moreover, we hypothesize that altered Treg homeostasis is a causative event for impaired resolution of inflammation and injury. The Candidate's recent and current work has provided her with the opportunity to develop her own research program and begin her transition to independence. The Career Development Plan described in the proposal outlines 2-years of mentored training with includes technical skills training in addition to career development activities designed to promote the successful transition to independence. A 3-year program of independent scientific and career development after successful recruitment as an Assistant Professor position is also outlined. The Candidate's Mentor has a proven track- record of excellent scientific productivity and successful mentorship and can provide the Candidate with a solid research environment in her lab at the Lerner Research Institute at the Cleveland Clinic. Research plan: Alcoholic liver disease (ALD) is remains a major socioeconomic burden. For decades, rates of morbidity and mortality have remained constant and the long-term prognosis for patients remains poor. ALD is multifactorial, mediated in part by the innate immune system. Complement, a component of innate immunity, is also implicated in the development of ALD and is becoming an attractive therapeutic target. Our early studies have identified that aberrant, uncontrolled complement activation contributes to tissue injury; however, complement is also required for liver healing. These studies provided the foundation for our current proposal: cell-specific complement therapies are a required alternative to current inhibitory therapies, which can be used as a novel treatment for ALD. Regulatory T cells (Treg) are key contributors to the resolution phase of inflammation and maintain immunological tolerance via secretion of a myriad of immunosuppressive and pro-resolving cytokines. Peripheral Treg are reduced in patients with advanced ALD, but the mechanism by which ethanol alters T cell homeostasis has not been investigated.  It is our working hypothesis that sustained complement activation due to ethanol leads to abnormal regulation of Treg in the liver, thus contributing to liver inflammation and tissue injury. In three specific aims, we will characterize ethanol-mediated Treg differentiation in the liver following Gao-Binge feeding in mice, explore the ability of adoptively transferred ex vivo differentiated iTreg to resolve liver inflammation and diminish liver injury in ethanol-fed mice, and assess the ability of extracellular vesicles to stimulate ex vivo differentiation of naïve CD4+ T cells to iTreg and Th17 subtypes. We expect the results of these aims will provide a strong foundation for future mechanistic studies and clinical interventions for ethanol- induced inflammation and tissue injury.