Kim-1 regulation of autophagy and MHC presentation in kidney injury

Kim-1 对肾损伤中自噬和 MHC 表达的调节

• 批准号: 8290478
• 负责人: Craig Robert Brooks
• 金额: $ 5.39万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8290478
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Address; Advanced Glycosylation End Products; Antigen Presentation; Antigens; Apoptotic; Autophagocytosis; Autophagosome; Bacteria; Cell Culture Techniques; Cell membrane; Cell model; Cells; Cessation of life; Chronic Kidney Failure; Cleaved cell; Clinical Trials; Cytoplasmic Tail; Data; Diagnosis; Drug Monitoring; Epithelial; Epithelial Cells; Exposure to; Extracellular Domain; Goals; Histocompatibility Antigens Class II; Human; Immune; In Vitro; Incidence; Inflammation; Injury; Kidney; Kidney Diseases; Kidney Failure; Lead; Left; Length; Life; Major Histocompatibility Complex; Mediating; Metalloproteases; Methods; Morbidity - disease rate; Mus; Necrosis; Pathway interactions; Patients; Peptides; Phagocytes; Phagocytosis; Phagocytosis Inhibition; Phagosomes; Play; Process; Proteins; Proximal Kidney Tubules; Qualifying; Regulation; Renal tubule structure; Research; Rodent; Role; Solutions; T-Cell Activation; T-Lymphocyte; Techniques; Testing; Therapeutic Intervention; Transmembrane Domain; Work; antigen processing; case-by-case basis; cell type; cellular imaging; chemical genetics; in vivo; injured; mortality; mutant; novel; oxidized low density lipoprotein; phosphatidylserine receptor; preclinical study; public health relevance; rat KIM-1 protein; scavenger receptor; trafficking; uptake

DESCRIPTION (provided by applicant): The overarching goal of the proposed work is to better understand the role of Kim-1 mediated phagocytosis in kidney injury. The long term goal is to identify potential regulatory mechanisms mediated by Kim-1 which could be used as targets to ameliorate kidney injury. Specifically, we will use in vitro and in vivo techniques to delineate the mechanism by which Kim-1 regulates autophagy and the effect Kim-1 mediated autophagy has on MHC class II presentation. In our first specific aim, the goal is to identify the mechanism by which Kim-1 upregulates autophagy and targets phagocytosed material to the autophagosome. Using chemical and genetic methods we will examine the trafficking of Kim-1 positive phagosomes to the autophagosome. We will also determine whether the cytoplasmic tail of Kim-1 targets phagocytosed debris to the autophagosome. Finally, we will examine whether Kim-1 cytoplasmic tail targets the phagosome to the autophagosome. In the second specific aim, we will examine whether Kim-1 induced autophagy leads to MHC class II presentation of peptides from the phagocytosed debris. We will approach this using two models cells not expressing Kim-1, expressing Kim-1 or expressing a phagocytosis null mutant and wt mice or mice expressing a phagocytosis null Kim- 1. First, we will co-localize Kim-1 with the autophagosome and MHC antigen processing compartment in cell culture and injured kidneys. We will then compare the presentation of peptides on MHC class II, following exposure to phagocytic targets. Finally, we will examine whether autophagy is necessary for Kim-1 expressing tubule cells to activate T-cells following phagocytosis. Relevance: Acute kidney injury remains a major kidney disease associated with high morbidity and mortality. The incidence of AKI has been steadily increasing over the past decade and is expected to increase further. The mortality rate of patients diagnosed with kidney failure is often 30-50%. Although research has lead to many treatment options and increased survivability, more research is needed to find a solution to this problem. Our study is aimed at better understanding the function of a protein (Kim-1) which is found associated with most forms of kidney failure. We will determine whether Kim-1 contributes to the autophagy and MHC presentation observed in kidney injury as well as identify potential targets for therapeutic intervention. PUBLIC HEALTH RELEVANCE: Acute kidney injury remains a major kidney disease associated with high morbidity and mortality. The incidence of AKI has been steadily increasing over the past decade and is expected to increase further. The mortality rate of patients diagnosed with kidney failure is often 30-50%. Although research has lead to many treatment options and increased survivability, more research is needed to find a solution to this problem. Our study is aimed at better understanding the function of a protein (Kim-1) which is found associated with most forms of kidney failure. We will determine how Kim-1 regulates autophagy and whether this autophagy contributes to the inflammation observed in kidney injury as well as identifies potential targets for therapeutic intervention.

描述(由申请人提供)：拟议工作的主要目标是更好地了解Kim-1介导的吞噬作用在肾脏损伤中的作用。长期目标是确定由Kim-1介导的潜在调控机制，这些机制可以作为减轻肾脏损伤的靶点。具体地说，我们将使用体外和体内技术来描述Kim-1调节自噬的机制以及Kim-1介导的自噬对MHC II类递呈的影响。在我们的第一个特定目标中，目标是确定Kim-1上调自噬并将吞噬的物质靶向自噬小体的机制。利用化学和遗传方法，我们将研究KIM-1阳性吞噬小体向自噬小体的运输。我们还将确定Kim-1的细胞质尾巴是否将吞噬的碎片靶向自噬小体。最后，我们将检查Kim-1细胞质Tail是否将吞噬小体靶向自噬小体。在第二个特定目标中，我们将检查Kim-1诱导的自噬是否导致吞噬的碎片中的多肽呈现MHC II类。我们将使用两个不表达Kim-1、表达Kim-1或表达吞噬功能缺失突变体的模型细胞和表达吞噬功能缺失Kim-1的小鼠或小鼠来探讨这一问题。首先，我们将在细胞培养和受损肾脏中将Kim-1与自噬小体和MHC抗原处理舱共定位。然后，我们将比较暴露于吞噬靶点后，MHC II类分子上的多肽呈现情况。最后，我们将检查在吞噬后，表达Kim-1的小管细胞是否需要自噬来激活T细胞。相关性：急性肾损伤仍然是一种与高发病率和死亡率相关的主要肾脏疾病。在过去的十年中，AKI的发病率一直在稳步上升，预计还会进一步增加。被诊断为肾衰竭的患者的死亡率通常为30%-50%。尽管研究已经导致了许多治疗选择和更高的存活率，但还需要更多的研究来找到这个问题的解决方案。我们的研究旨在更好地了解一种蛋白质(Kim-1)的功能，这种蛋白质被发现与大多数形式的肾衰竭有关。我们将确定Kim-1是否与肾脏损伤中观察到的自噬和MHC呈现有关，并确定潜在的治疗干预靶点。 公共卫生相关性：急性肾损伤仍然是一种与高发病率和死亡率相关的主要肾脏疾病。在过去的十年中，AKI的发病率一直在稳步上升，预计还会进一步增加。被诊断为肾衰竭的患者的死亡率通常为30%-50%。尽管研究已经导致了许多治疗选择和更高的存活率，但还需要更多的研究来找到这个问题的解决方案。我们的研究旨在更好地了解一种蛋白质(Kim-1)的功能，这种蛋白质被发现与大多数形式的肾衰竭有关。我们将确定Kim-1如何调节自噬，以及这种自噬是否有助于肾脏损伤中观察到的炎症，并确定潜在的治疗干预靶点。