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

• 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
• 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

 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（通过应用提供）：肝脏居民巨噬细胞的激活和促炎性细胞因子产生的增加是酒精性肝病（ALD）发病机理的标志。酒精诱导的氧化应激在巨噬细胞激活中起重要作用。应力诱导的热休克蛋白（HSP）充当信号分子的分子伴侣，对巨噬细胞激活很重要。热休克蛋白90（HSP90）及其ER形式GRP94/GP96是TLR4途径中信号分子的关键链酮，因此调节炎症细胞因子。 HSP90和GP96一起可以连接应力和炎症途径，并在ALD的发展中发挥重要作用。该项目的最后一个资金期间的研究确定：1）HSP90α（细胞质）和GP96/GRP94（HSP90的内质网（ER）形式），但HSP70在肝型巨噬细胞中的增加而不是HSP70在肝果实中的增加，以及人类含量的小鼠和人类酒精含量的IRF 3的hepatip90/2）hsp90/2）促进TLR4信号传导2）使用药物17-DMAG保护小鼠免受酒精性肝损伤的抑制作用，3）髓样特异性GP96缺乏减轻酒精肝损伤4）HSF1，尽管通过酒精激活了HSP90α的诱导。另一方面，HSP90抑制剂在减少损伤过程中诱导的HSF1和HSP70对于减少肝脏炎症反应至关重要。在这里，我们假设细胞质HSP90α和ER GP96/GRP94有助于肝巨噬细胞激活，并且是ALD中合理的治疗靶标。增加的Hsp90α促进巨噬细胞激活，而GP96（未折叠的蛋白质反应（UPR）基因）诱导的Toll样受体4表达表达，并且反应性过高有助于酒精性肝病中的巨噬细胞敏感性。最后，我们建议HSF1和HSP70在HSP90抑制剂期间发挥抗炎作用，将有助于解开应激蛋白和炎症反应之间交叉访问的创新途径，并将HSP90作为ALD中的新治疗靶标建立。具体目的如下：1）研究ALD中HSP90α抑制对巨噬细胞激活的影响： b）检查HSP90α抑制对肝脏巨噬细胞极化的影响。 c）分析巨噬细胞特异性HSP90α抑制对酒精脂肪变性的旁分泌作用。 2）研究HSP90的髓样GP96的功能，在酒精诱导的巨噬细胞敏感性中通过：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。 C）表征GP96在ALD期间巨噬细胞UPR激活中的作用。 3）评估ALD中HSP90抑制期间HSF1和HSP70的抗炎作用：a）评估髓样特异性HSF1在巨噬细胞激活和ALD中的作用。 b）研究HSF1在17-DMAG处理和抑制ALD中的作用。 c）确定Hsp70在17-DMAG处理过程中的抗炎反应中的作用。