Heat shock protein 90 in alcoholic liver disease: targeting macrophage function

酒精性肝病中的热休克蛋白 90：针对巨噬细胞功能

基本信息

批准号：
9302598
负责人：
Pranoti Mandrekar
金额：
$ 37.51万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9302598
关键词：
Affect Alcohol consumption Alcoholic Hepatitis Alcoholic Liver Diseases Alcohols Anti-Inflammatory Agents Anti-inflammatory Chronic Cirrhosis Clinical Development Disease Drug Delivery Systems Drug Targeting Endoplasmic Reticulum Endotoxins Fatty Liver Funding Gene Targeting Genes HSF1 HSP 90 inhibition Heat Stress Disorders Heat shock proteins Heat-Shock Proteins 90 Hepatic Hepatocyte Human IRF3 gene Immune Cell Activation Inflammation Inflammatory Inflammatory Response Injury Kupffer Cells Link Liver Macrophage Activation Malignant Neoplasms Mediating Molecular Chaperones Morbidity - disease rate Mus Myelogenous Oxidative Stress Pathogenesis Pathway interactions Pharmacology Play Primary carcinoma of the liver cells Production Property Proteins Regulation Reporting Resolution Role Signal Pathway Signal Transduction Signaling Molecule Small Interfering RNA Steatohepatitis Stimulus Stress Surface TLR4 gene Therapeutic base biological adaptation to stress cell injury cytokine global health in vivo inhibitor/antagonist innovation lipid metabolism liver injury macrophage mortality nanoemulsion nanoparticle new therapeutic target novel novel therapeutics paracrine paralogous gene pre-clinical problem drinker public health relevance response stress protein therapeutic target transcription factor

项目摘要

DESCRIPTION (provided by applicant): Activation of liver resident macrophages and increased pro-inflammatory cytokine production is a hallmark in the pathogenesis of alcoholic liver disease (ALD). Alcohol-induced oxidative stress plays an important role in macrophage activation. Stress induced heat shock proteins (hsps) function as molecular chaperones of signaling molecules important in macrophage activation. Heat shock protein 90 (hsp90) and its ER form, Grp94/gp96 are crucial chaperones of signaling molecules in the TLR4 pathway and thus regulate inflammatory cytokines. Together, hsp90 and gp96 can link stress and inflammatory pathways and play an important role in development of ALD. Studies during the last funding period of this project established that: 1) hsp90α (cytoplasmic) and gp96/grp94 (endoplasmic reticulum (ER) form of hsp90), but not hsp70 is increased in hepatic macrophages and whole livers of chronic alcohol-fed mice and in human alcoholic hepatitis livers, 2)Hsp90α chaperones IKKβ and IRF3 and facilitates TLR4 signaling 2) Inhibition of hsp90 in vivo using pharmacological agent 17-DMAG protects mice from alcoholic liver injury, 3) Myeloid-specific gp96 deficiency alleviates alcoholic liver injury 4) HSF1, although activated by alcohol is not sufficient for induction of hsp90α. On the other hand, HSF1 and hsp70 induced by hsp90 inhibitors during decreased injury is crucial for reduction of liver inflammatory responses. Here we hypothesize that cytoplasmic hsp90α and ER gp96/grp94 contribute to hepatic macrophage activation and are plausible therapeutic targets in ALD. Increased hsp90α promotes macrophage activation and gp96 (unfolded protein response (UPR) gene) induced toll-like receptor 4 surface expression and hyperresponsiveness contributes to macrophage sensitization in alcoholic liver disease. Finally we propose that HSF1 and hsp70 exert anti-inflammatory effects during hsp90 inhibition in ALD. Use of hsp90 inhibitors will help unravel innovative pathways of cross-talk between stress proteins and inflammatory responses and establish hsp90 as a new therapeutic target in ALD. The Specific Aims are as follows: 1) To study the effect of hsp90α inhibition on macrophage activation in ALD by: A) Using hsp90α siRNA to target liver macrophages and 17-DMAG packaged nanoparticles in ALD. B) Examining the effect of hsp90α inhibition on macrophage polarization in the liver. C) Analysis of paracrine effect of macrophage specific hsp90α inhibition on alcoholic steatosis. 2) To investigate the function of myeloid gp96, ER paralog of hsp90, in alcohol-induced sensitization of macrophages by: A) Determine the regulation of gp96 expression during alcoholic liver injury. B) Study importance of gp96/TLR4 interaction and TLR4 hyperresponsiveness during ALD. C) Characterize the role of gp96 in macrophage UPR activation during ALD. 3) To assess anti-inflammatory effects of HSF1 and hsp70 during hsp90 inhibition in ALD: A) Evaluate the role of myeloid-specific HSF1 in macrophage activation and ALD. B) Investigate the role of HSF1 during 17-DMAG treatment and inhibition of ALD. C) Determine the role of hsp70, in anti-inflammatory responses during 17- DMAG treatment.

描述（由申请人提供）：驻留肝脏巨噬细胞的激活和促炎细胞因子的产生增加是酒精性肝病（ALD）发病机制的一个标志。酒精诱导的氧化应激在应激诱导的热休克中发挥重要作用。蛋白 (hsps) 作为信号分子的分子伴侣，在巨噬细胞激活中发挥重要作用，热休克蛋白 90 (hsp90) 及其 ER 形式， Grp94/gp96 是 TLR4 通路中信号分子的关键伴侣，因此可以共同调节炎症细胞因子，从而将应激和炎症通路联系起来，并在该项目最后资助期间的 ALD 研究中发挥重要作用。 1) hsp90α（细胞质）和 gp96/grp94（hsp90 的内质网 (ER) 形式），但是慢性酒精喂养小鼠的肝巨噬细胞和整个肝脏以及人类酒精性肝炎肝脏中的 hsp70 增加，2)Hsp90α 分子伴侣 IKKβ 和 IRF3 并促进 TLR4 信号传导 2) 使用药剂 17-DMAG 体内抑制 hsp90 可保护小鼠免受酒精性肝损伤，3) 骨髓特异性 gp96 缺乏可减轻酒精性肝损伤 4) HSF1 虽然被酒精激活不足以诱导 hsp90α，但在减少损伤期间，hsp90 抑制剂诱导的 HSF1 和 hsp70 对于减少肝脏炎症反应至关重要。 hsp90α 的增加可促进巨噬细胞活化，并且是 ALD 中可能的治疗靶标。最后，我们提出，HSF1 和 hsp70 在 ALD 中的 hsp90 抑制过程中发挥抗炎作用，这将有助于使用 hsp90 抑制剂。揭示应激蛋白与炎症反应之间相互作用的创新途径，并将 hsp90 确立为 ALD 的新治疗靶点。如下：1) 通过以下方式研究 hsp90α 抑制对 ALD 中巨噬细胞活化的影响：A) 使用 hsp90α siRNA 靶向 ALD 中的肝脏巨噬细胞和 17-DMAG 包装的纳米颗粒 B) 检查 hsp90α 抑制对肝脏中巨噬细胞极化的影响。 C)巨噬细胞特异性hsp90α抑制对酒精性脂肪变性的旁分泌作用分析2)研究其功能。骨髓 gp96（hsp90 的 ER 旁系同源物）在酒精诱导的巨噬细胞致敏中的作用：A) 确定酒精性肝损伤期间 gp96 表达的调节 B) 研究 ALD 期间 gp96/TLR4 相互作用和 TLR4 高反应性的重要性。 ALD 期间 gp96 在巨噬细胞 UPR 激活中的作用 3) 评估 HSF1 和 HSF1 的抗炎作用。 hsp70 在 ALD 中 hsp90 抑制过程中的作用：A) 评估骨髓特异性 HSF1 在巨噬细胞激活和 ALD 中的作用 B) 研究 HSF1 在 17-DMAG 治疗和抑制 ALD 过程中的作用 C) 确定 hsp70 在抗 ALD 中的作用。 -17-DMAG治疗期间的炎症反应。