Role of necroptosis in inflammation and NAFLD to HCC progression

坏死性凋亡在炎症和 NAFLD 与 HCC 进展中的作用

• 批准号: 10580847
• 负责人: Deepa Sathyaseelan
• 金额: $ 7.25万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10580847
• 关键词: Acceleration; Affect; Apoptosis; Binding; Cancer Etiology; Cell Membrane Permeability; Cell Surface Receptors; Cells; Cessation of life; Choline Deficiency; Chronic; Cirrhosis; Data; Development; Diet; Diethylnitrosamine; Disease; Fibrosis; Genetic; HMGB1 gene; Hepatic; Hepatocyte; High Fat Diet; Immune; Immunologic Receptors; Incidence; Individual; Infiltration; Inflammation; Knock-in Mouse; Knockout Mice; Knowledge; Liver; Malignant neoplasm of liver; Measurement; Mediating; Mediator; Membrane; Molecular; Mus; Obesity; Obesity Epidemic; Pathway interactions; Patients; Pattern; Phosphorylation; Phosphotransferases; Play; Population; Primary carcinoma of the liver cells; Protein-Serine-Threonine Kinases; Proteins; RIPK3 gene; Research; Risk Factors; Role; Serine; Source; Testing; United States; chronic liver disease; diet-induced obesity; feeding; genetic approach; inhibitor; liver inflammation; mouse model; non-alcoholic fatty liver; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; obese person; obesity prevention; overexpression; pharmacologic; prevent; receptor; response; superoxide dismutase 1; systemic inflammatory response

Project Summary/Abstract The research objective is to identify the source of chronic inflammation in obesity-mediated hepatocellular carcinoma (HCC). Hepatocellular carcinoma (HCC), a major form of liver cancer, is the fourth leading cause of cancer-related deaths worldwide. In recent years, obesity has emerged as the major and independent risk factor for HCC, and HCC is predicted to the third leading cause of cancer-related deaths in the United States by 2030 due to obesity epidemic. Obesity-induced nonalcoholic fatty liver disease (NAFLD), which affects nearly 25% of the US population, is a key driver of HCC in obese individuals. Despite this strong association between obesity and HCC, the mechanisms that drive HCC development in obesity is not clearly understood. Non-resolving chronic inflammation is a major contributor to the development and progression of HCC in obesity and damage associated molecular patterns (DAMPs) are one of the proposed mediators of HCC. Necroptosis is a programmed cell death that has been shown to play a major role in inflammation through the release of DAMPs, which bind to cell surface receptors of immune cells to induce inflammation. In this application, we propose that hepatocyte necroptosis in NAFLD is a key factor in HCC progression. Our preliminary data support the role of necroptosis in HCC, e.g., blocking necroptosis using Ripk3-/- or Mlkl-/- mice reduced hepatic inflammation, infiltration of innate immune cells, and HCC incidence in mice fed a choline deficient-high fat diet (CD-HFD) that do not induce obesity. In a genetic mouse model of spontaneous HCC (Sod1-/- mice) inhibiting necroptosis using necroptosis inhibitor, necrostatin-1s, reduce hepatic necroptosis, inflammation, and pathways mediating HCC development. Based on this, we propose that hepatocyte necroptosis is the major source of hepatic inflammation in the microenvironment where HCC is initiated. We hypothesize that progression of NAFLD to HCC is due to increased inflammation that arises from hepatocyte necroptosis and that preventing necroptosis will reduce inflammation and progression of NAFLD to HCC. This hypothesis will be tested using genetic approaches to block or activate necroptosis in hepatocytes by targeting RIPK3 or MLKL, kinases in the necroptosis pathway, and assessing its effect on inflammation and HCC. In Aim 1, necroptosis will be specifically reduced/blocked in hepatocytes using hepatocyte specific Ripk3 or Mlkl knockout mice, and determine if blocking hepatocyte necroptosis reduces inflammation and delays/reduces HCC development in response to an HCC- inducing diet; In Aim 2, necroptosis will be induced specifically in hepatocytes by overexpressing RIPK3 or MLKL using a novel Ripk3 or Mlkl knockin mouse model we have developed to determine if inducing necroptosis in hepatocytes leads to an increase in liver inflammation and increased progression and incidence of HCC.

项目概要/摘要 研究目的是确定肥胖介导的肝细胞慢性炎症的根源 癌（HCC）。肝细胞癌 (HCC) 是肝癌的一种主要形式，是导致肝癌的第四大原因。 全球癌症相关死亡。近年来，肥胖已成为主要且独立的危险因素 预计到 2030 年，HCC 将成为美国癌症相关死亡的第三大原因 由于肥胖流行。肥胖引起的非酒精性脂肪肝 (NAFLD) 影响近 25% 美国人口中，肥胖者是 HCC 的主要驱动因素。尽管肥胖与肥胖之间存在密切联系 和 HCC，肥胖导致 HCC 发展的机制尚不清楚。未解决 慢性炎症是肥胖和损害中 HCC 发生和进展的主要因素 相关分子模式 (DAMP) 是 HCC 的拟议介质之一。坏死性凋亡是一种 程序性细胞死亡已被证明通过释放 DAMP 在炎症中发挥重要作用， 它与免疫细胞的细胞表面受体结合以诱导炎症。在本申请中，我们建议 NAFLD 中的肝细胞坏死性凋亡是 HCC 进展的关键因素。我们的初步数据支持以下作用： HCC 中的坏死性凋亡，例如，使用 Ripk3-/- 或 Mlkl-/- 小鼠阻断坏死性凋亡可减少肝脏炎症， 喂食缺乏胆碱的高脂肪饮食 (CD-HFD) 的小鼠的先天免疫细胞浸润和肝癌发生率 不会诱发肥胖。在自发性 HCC 基因小鼠模型（Sod1-/- 小鼠）中，使用抑制坏死性凋亡 坏死性凋亡抑制剂 necrostatin-1s 可减少肝坏死性凋亡、炎症和介导 HCC 的途径 发展。据此，我们提出肝细胞坏死性凋亡是肝脏炎症的主要根源。 发生 HCC 的微环境。我们假设 NAFLD 进展为 HCC 的原因是 肝细胞坏死性凋亡引起的炎症增加，预防坏死性凋亡将 减少炎症和 NAFLD 向 HCC 的进展。该假设将通过遗传来检验 通过靶向 RIPK3 或 MLKL（肝细胞中的激酶）来阻断或激活肝细胞坏死性凋亡的方法 坏死性凋亡途径，并评估其对炎症和 HCC 的影响。在目标 1 中，坏死性凋亡将具体是 使用肝细胞特异性 Ripk3 或 Mlkl 敲除小鼠在肝细胞中减少/阻断，并确定是否阻断 肝细胞坏死性凋亡可减少炎症并延迟/减少 HCC 的发展，以应对 HCC- 诱导饮食；在目标 2 中，通过过表达 RIPK3 或 MLKL 将在肝细胞中特异性诱导坏死性凋亡 使用我们开发的新型 Ripk3 或 Mlkl 敲入小鼠模型来确定是否诱导坏死性凋亡 肝细胞导致肝脏炎症增加，并增加肝癌的进展和发病率。