Role of MLKL in Alcohol-associated Liver Disease

MLKL 在酒精相关性肝病中的作用

• 批准号: 10889380
• 负责人: Xiaoqin Wu
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10889380
• 关键词: Acute; Alcohol-Induced Disorders; Alcoholic Hepatitis; Alcoholic Liver Diseases; Anti-Inflammatory Agents; Apoptotic; Bone Marrow; Bone Marrow Transplantation; Cell Culture Techniques; Cell Death; Cell Survival; Cell model; Cells; Cessation of life; Chimera organism; Chronic; Cirrhosis; Clinical; Communication; Complex; Data; Dedications; Development; Development Plans; Disease; Disease Progression; Economic Burden; Environment; Equilibrium; Ethanol; Exhibits; Exposure to; Foundations; Functional disorder; Future; Heavy Drinking; Hepatic; Hepatitis; Hepatocyte; Heterogeneity; Histone H3; Homeostasis; Human; Immune; Immune response; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injury; Intervention; Knock-out; Kupffer Cells; Liver; Liver diseases; Macrophage; Maintenance; Measures; Mediating; Mediator; Mentors; Mentorship; Modeling; Molecular; Morbidity - disease rate; Morphology; Mus; Myeloid Cells; Natural Immunity; Necrosis; Outcome; Paper; Pathogenesis; Pathway interactions; Patients; Peripheral; Phase; Phenotype; Phosphotransferases; Play; Population; Positioning Attribute; Primary carcinoma of the liver cells; Process; Productivity; RIPK1 gene; RIPK3 gene; Regulation; Research; Research Personnel; Resolution; Role; S100A8 gene; Signal Transduction; Solid; Stains; Technical Expertise; Testing; Therapeutic Intervention; Tissues; Training; United States; Work; alcohol exposure; alcohol response; career; career development; design; extracellular; in vivo; injury and repair; insight; liver inflammation; monocyte; mortality; mouse model; neutrophil; phosphoproteomics; prevent; professor; programs; recruit; response; response to injury; restoration; skill acquisition; socioeconomics; tissue injury

Project summary The Candidate is a young investigator dedicated to developing an academic career focused on understanding the mechanisms by which ethanol disrupts balance between immune cell survival and death pathways. With a strong background in liver diseases and related innate immunity, the candidate has developed particular expertise in the use of in vivo and cell culture models to conduct the proposed studies. The Candidate’s current work has provided her with the opportunity to develop her own research program and begin her transition to independence. The Career Development Plan outlines 2-years of mentored training on technical skills and 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. Research plan: Alcohol-associated liver disease (ALD) remains a major socioeconomic burden with high morbidity and mortality. Activation of innate immunity not only contributes to progression of ALD, but is also critical for resolution of injury. Appropriate regulation of pro-survival and pro-death pathways is critical to the ability of innate immune cells to rapidly respond to maintain liver homeostasis in ethanol-induced injury. MLKL-mediated necroptosis plays a dual role in tissue injury/repair. Whether MLKL-mediated necroptosis of immune cells is beneficial or detrimental in the context of ethanol is unknown. In preliminary work, Mlkl deficiency in myeloid cells exacerbated ethanol-induced injury, associated with increased ethanol-induced accumulation of hepatic macrophages and neutrophils. Importantly, we found that Gao-binge increased necrosis/necroptosis of hepatic F4/80+ cells, suggesting that MLKL may protect from ethanol-induced injury by promoting macrophage death and resolution of inflammation. Together, these data led us to hypothesize that myeloid cell-specific MLKL restricts ethanol- induced injury by regulating hepatic immune cell survival and death. To test this hypothesis, hepatic immune cell populations and death will be characterized by flow cytometric analysis of isolated NPCs from livers of Mlkl BM chimeras after Gao-binge. We will then challenge cell-specific knock-outs (LysM, Clec4f and MRP8 CRE crossed with Mlklfl/fl) to ethanol, to distinguish cell-specific contributions of MLKL to mALD. Further, a phospho-proteomics paired with targeted mechanistic approaches will be utilized to determine whether ethanol-mediated activation of MLKL is RIP3-dependent and/or RIP3-independent. Additionally, since neutrophils and neutrophil extracellular traps (NETs) are important in ALD, the project will further explore whether neutrophil-specific MLKL limits ethanol-induced injury by regulating NETs. Altogether, this proposal will provide a solid foundation for future mechanistic studies and clinical interventions for ethanol-induced injury and inflammation.

项目摘要 候选人是一名年轻的研究人员，致力于发展学术生涯，专注于理解 乙醇破坏免疫细胞存活和死亡途径之间平衡的机制。与 在肝脏疾病和相关先天免疫方面有很强的背景， 使用体内和细胞培养模型进行拟议研究的专业知识。候选人目前 工作为她提供了发展自己的研究计划的机会，并开始向 独立职业发展计划概述了2年的技术技能指导培训， 职业发展活动，旨在促进成功过渡到独立。3年计划 独立的科学和职业发展后，成功招聘为助理教授职位 还概述了。候选人的导师具有良好的科学生产力的良好记录， 成功的指导，并能在她的实验室为候选人提供一个坚实的研究环境。研究 计划：酒精相关性肝病（ALD）仍然是一个主要的社会经济负担，发病率高， mortality.先天免疫的激活不仅有助于ALD的进展，而且对于解决ALD也至关重要。 伤害。适当调节促存活和促死亡途径对于先天免疫的能力至关重要。 细胞迅速响应，以维持乙醇诱导的损伤中的肝脏稳态。MLKL介导的坏死性凋亡作用 在组织损伤/修复中的双重作用。MLKL介导的免疫细胞坏死性凋亡是否有益， 在乙醇的情况下是有害的是未知的。在前期工作中，骨髓细胞中的Mlkl缺乏加剧了 乙醇诱导的损伤，与乙醇诱导的肝巨噬细胞蓄积增加相关， 中性粒细胞重要的是，我们发现高斌格增加肝F4/80+细胞的坏死/坏死性凋亡， 提示MLKL可能通过促进巨噬细胞死亡和消退来保护免受乙醇诱导的损伤 炎症。总之，这些数据使我们假设骨髓细胞特异性MLKL限制乙醇- 通过调节肝脏免疫细胞的存活和死亡诱导损伤。为了验证这一假设， 将通过对来自Mlkl BM的肝的分离的NPC的流式细胞术分析来表征群体和死亡。 高宾哥之后的嵌合体然后，我们将挑战细胞特异性敲除（LysM、Clec 4f和MRP 8 CRE杂交）。 用Mlklfl/fl）与乙醇的比较，以区分MLKL对mALD的细胞特异性贡献。此外，磷酸蛋白质组学 与靶向机制方法配对将用于确定乙醇介导的激活是否 是RIP 3依赖的和/或RIP 3独立的。此外，由于中性粒细胞和中性粒细胞外 陷阱（NET）在ALD中很重要，该项目将进一步探讨是否有嗜酸性粒细胞特异性MLKL限制 乙醇诱导的损伤通过调节NET。总的来说，这项建议将为今后的工作奠定坚实的基础。 乙醇诱导的损伤和炎症的机制研究和临床干预。