Role of Intestinal Proteostasis mediator HSP90 in alcoholic liver disease.

肠道蛋白质稳态介质 HSP90 在酒精性肝病中的作用。

基本信息

批准号：
10493334
负责人：
Pranoti Mandrekar
金额：
$ 19.89万
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：
2021
资助国家：
美国
起止时间：
2021-09-25 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10493334
关键词：
Affect Agonist Alcohol abuse Alcoholic Liver Diseases Alcohols Antibiotics Bacteria Bacteriophages Blood Circulation Cell Adhesion Molecules Cells Chronic Client Colitis Colon Carcinoma Data Disease Economic Burden Encapsulated Endoplasmic Reticulum Endotoxins Epithelial Exhibits Foundations Functional disorder Future Goals Health Heat shock proteins Heat-Shock Proteins 90 IRAK1 gene Immune Immunologic Receptors Impairment Inflammation Inflammatory Inflammatory Response Injury Intestines Lamina Propria Leaky Gut Liver diseases MAP Kinase Gene Mammals Mediating Mediator of activation protein Messenger RNA Molecular Chaperones Molecular Target Mus Myelogenous Nutrient Organism Pathogenicity Pathway interactions Permeability Pharmacology Phenotype Play Post-Translational Protein Processing Probiotics Property Protein Isoforms Proteins Proteome Reporting Role Signal Pathway Signaling Molecule Small Intestines Stress TNF gene Tight Junctions Toll-like receptors Virus alcohol exposure base cytokine deprivation drug development dysbiosis fecal transplantation fungus gut homeostasis gut inflammation gut microbiome gut microbiota ileum immune activation in vivo inhibitor intercellular cell adhesion molecule intestinal barrier jejunum liver inflammation macrophage microbial microbiome microbiota nano nanoparticle paralogous gene problem drinker proteostasis response targeted treatment therapeutic target

项目摘要

ABSTRACT Chronic alcohol alters the enteric microbiome (dysbiosis), induces inflammation, impairs the gut barrier function resulting in a “leaky” gut, passage of microbial products into portal circulation and eventually liver inflammation. Current therapies targeting the microbiome or host pathways to restore barrier function have utilized antibiotics, probiotics, synbiotics, fecal microbiome transplant (FMT), bacteriophage therapy, FXR agonists and nutrient- based treatments. The major goal of this proposal is to investigate the pathogenic role of proteostasis mediator, HSP90 in the gut by intestinal targeting in vivo using specific pharmacological inhibitors to evaluate its effect on alcohol mediated host intestinal inflammation, barrier function and dysbiosis. Two predominant forms of mammalian stress inducible HSP90 paralogs are cytoplasmic (CYT) HSP90AA1 and HSP90B1/GP96 in the endoplasmic reticulum (ER). The chaperone function of HSP90 and GP96 is crucial in fine tuning immune cell activation and inflammatory responses. The precise role of proteostasis chaperones such as HSP90 in chronic alcohol mediated intestinal dysfunction is not yet investigated. We reported that targeting HSP90 using specific pharmacological inhibitor 17-DMAG, reduced gut derived circulating endotoxin in ALD. Myeloid specific HSP90B1/GP96 deficient mice also exhibit reduced circulating endotoxin in ALD. Further our preliminary data show induction of HSP90AA1 and HSP90B1/GP96 mRNA in intestine of chronic alcohol exposed mice. In addition, 17-DMAG is bioavailable in the intestine and inhibits inflammatory cytokine, TNFα in the jejunum and ileum. We hypothesize that chronic alcohol induces HSP90 in host intestine contributing to inflammation and compromising gut barrier function in ALD. In addition, since therapeutic targeting of gut inflammation can restore protective microbiota, as well as considering the antibiotic and antimycotic properties of 17-DMAG, it is likely that 17-DMAG treatment favors protective gut microbiota, facilitated by reduced host intestinal inflammation. Collectively, here we will explore the role of HSP90 in alcohol mediated gut inflammation and barrier function and also assess whether pharmacological targeting of host HSP90 impacts gut microbiota. The Specific Aims are: 1) To delineate the role of HSP90 isoforms on alcohol induced gut inflammation by- Evaluating expression of HSP90 isoforms, intestinal targeting of HSP90 using 17-DMAG encapsulated nanoparticles to study inflammatory responses in small intestine and immune cells in lamina propria and assessing whether myeloid specific HSP90B1/GP96 deficiency affects inflammatory responses in the gut. 2) To assess the functional relevance of HSP90 on alcohol mediated intestinal barrier integrity and dysbiosis by - Evaluating permeability and epithelial tight junction proteins during HSP90 and myeloid GP96 inhibition. B) Assessing the effect of intestine targeted nano-DMAG and myeloid GP96 deficiency on microbiota. Successful completion of the proposed studies will uncover the role of proteostasis chaperones in the gut and guide future studies to identify proteostasis pathways in alcohol mediated gut injury.

抽象的慢性酒精改变肠性微生物组（营养不良），诱发感染，损害肠道障碍功能导致肠道“漏水”，将微生物产物通过门户循环，并最终注射肝脏。针对微生物组或宿主途径恢复屏障功能的当前疗法已使用抗生素，益生菌，合成生物，粪便微生物组移植（FMT），噬菌体疗法，FXR激动剂和营养 - 基于治疗。该提案的主要目的是研究蛋白质质介质的致病作用，肠道中的Hsp90通过使用特定的药物抑制剂在体内进行肠道靶向肠道靶向酒精介导的宿主肠道感染，屏障功能和营养不良。两种主要形式哺乳动物应力诱导HSP90旁系同源物是细胞质（CYT）HSP90AA1和HSP90B1/GP96 内质网（ER）。 HSP90和GP96的伴侣函数对于微调免疫球至关重要激活和炎症反应。 Proteostasis伴侣（例如HSP90）在慢性中的精确作用酒精介导的肠道功能障碍尚未研究。我们报道了使用特定的针对HSP90的药理学抑制剂17-DMAG，降低了ALD中的肠道循环内毒素。髓样特异性 HSP90B1/GP96不足的小鼠还暴露了ALD中循环内毒素的减少。进一步我们的初步数据显示慢性酒精暴露小鼠肠中HSP90AA1和HSP90B1/GP96 mRNA的诱导。在此外，17-DMAG在肠中可生物利用，并抑制炎症性细胞因子，空肠中的TNFα和回肠。我们假设慢性酒精会在宿主肠道中诱导HSP90，从而导致炎症和 ALD中损害肠道屏障功能。另外，由于肠道注射的治疗靶向可以恢复保护性微生物群，以及考虑17-DMAG的抗生素和抗菌特性，很可能 17-DMAG治疗有利于通过减少宿主肠道注射制备的保护性肠道菌群。总体而言，我们将在这里探索HSP90在酒精介导的肠道注射和屏障功能中的作用还评估宿主HSP90的药理靶向是否会影响肠道菌群。具体目标为：1）描绘Hsp90同工型在饮酒诱导的肠道注射中的作用，并评估表达 HSP90同工型，使用17-DMAG封装的纳米颗粒对HSP90进行肠道靶向小肠和免疫细胞的炎症反应在固有层中，并评估髓样特定的HSP90B1/GP96缺乏会影响肠道中的炎症反应。 2）评估功能 HSP90对酒精介导的肠道屏障完整性和营养不良的相关性 - 通过评估渗透性 HSP90和髓样GP96期间的上皮紧密连接蛋白。 b）评估的影响肠道靶向纳米-DMAG和髓样GP96在微生物群上缺乏。成功完成拟议的研究将揭示蛋白结论性伴侣在肠道中的作用，并指导未来的研究以识别酒精介导的肠道损伤中的蛋白质级途径。