The Role of the IL-1 Receptor in the AKI to CKD transition

IL-1 受体在 AKI 向 CKD 转变中的作用

• 批准号: 10370332
• 负责人: Jamie R Privratsky
• 金额: $ 17.8万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370332
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Animal Model; Anti-Inflammatory Agents; Applications Grants; Area; Attenuated; Behavior; Binding; CD4 Positive T Lymphocytes; Cell Communication; Cell Death; Cell Surface Receptors; Cell physiology; Cell surface; Cells; Chronic; Chronic Kidney Failure; Coculture Techniques; Complex; Complication; Critical Care; Critical Illness; Cytometry; Death Rate; Dendritic Cells; Development; Fibrosis; Foundations; Functional disorder; Future; Gene Activation; Gene Expression; Goals; Health; Health Care Costs; Human; Immune; Immune signaling; Immune system; In Vitro; Inflammatory; Injury; Injury to Kidney; Innate Immune Response; Interleukin-1 Receptors; Interleukins; Ischemia; Kidney; Knowledge; Leukocytes; Life; Lymphocyte; Lymphocyte Activation; Measures; Mediating; Mediator of activation protein; Medicine; Methods; Mission; Modeling; Morbidity - disease rate; Multiple Organ Failure; Mus; Myeloid Cells; Nature; Operative Surgical Procedures; Organ failure; Organism; Patients; Phenotype; Play; Population; Process; Production; Public Health; Quality of life; Receptor Activation; Receptor Signaling; Recovery; Regulatory T-Lymphocyte; Reperfusion Therapy; Research; Research Personnel; Role; Sepsis; Severities; Signal Transduction; Spatial Distribution; Syndrome; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Techniques; Testing; Therapeutic; Tissues; Toxin; Training; Tubular formation; United States National Institutes of Health; Ureteral obstruction; Work; adaptive immune response; base; body system; cell injury; cell motility; cytokine; disability; draining lymph node; experience; healing; immunomodulatory therapies; immunoregulation; improved; kidney cell; kidney fibrosis; lymph nodes; macrophage; migration; mortality; next generation sequencing; novel; novel imaging technique; organ repair; prevent; programs; renal damage; repaired; septic; single-cell RNA sequencing; therapy design; tissue injury

ABSTRACT Multi-organ dysfunction syndrome (MODS) leads to significant morbidity and mortality in critically ill patients. Acute kidney injury (AKI) is one of the most common components of MODS. Unresolved AKI leads to ongoing renal injury, fibrosis, and subsequent chronic kidney disease (CKD). However, the mechanisms that direct renal recovery and prevent the AKI to CKD transition are poorly understood. One novel and promising therapeutic approach is to modulate the functions of macrophages and dendritic cells (DCs) that accumulate in the injured kidney in order to prevent further injury and fibrosis. Interleukin (IL)-1α/β are canonical pro-inflammatory cytokines that activate macrophages and DCs after binding to their cell surface receptor, IL-1R1. IL-1R1 activation plays a prominent role in renal healing. However, IL-1R1’s specific effects on macrophage and DC function following AKI and on the AKI to CKD transition are poorly understood. Based on our preliminary studies, our central hypothesis is that following AKI, macrophage IL-1R1 activation triggers TNFα (TNF)-dependent renal cell necroptosis, which directs IL-1R1-dependent migration of DCs to lymph nodes to activate pro-inflammatory T cells to drive the AKI to CKD transition. We will test our central hypothesis as follows: Specific Aim 1: Determine effects of macrophage IL-1R1 activation on TNF-mediated renal cell necroptosis following ischemic AKI. Mice with macrophage-specific deletion of IL-1R1 (IL-1R1 MKO) and controls (IL-1R1 MWT) will undergo ischemia/reperfusion (I/R)- and septic AKI, and the severity of acute kidney damage will be quantified. Renal cell necroptosis will be measured in injured kidneys and in macrophage-renal tubular cell (RTC) co-culture. We will employ cell surface phenotyping and single cell RNA-seq on sorted leukocytes from injured kidneys to precisely phenotype myeloid cells to determine how IL-1R1 activation modulates macrophage polarization and renal injury. Specific Aim 2: Elucidate the role of IL-1R1-mediated dendritic cell migration and T cell activation during the AKI to CKD transition. Mice with deletion of IL-1R1 in DCs (IL-1R1 DCKO) and controls (IL-1R1 DCWT) will be subjected to I/R-induced AKI, and at multiple timepoints, the severity of renal damage and fibrosis will be compared. We will examine IL-1R1-mediated accumulation of DC subsets in renal lymph nodes following I/R and their consequent activation of effector lymphocytes. We will use novel tissue cytometry to analyze spatial distribution of DC and T cells within fibrotic areas. Activated DC and T cells will be co-cultured with RTC to determine their effect on pro-fibrotic gene expression programs. The proposed studies will have a significant positive impact by promoting the development of future targeted treatments for patients with life-threatening AKI. Together with my training plan, these studies will lay the foundation for my development as an independent investigator in critical care medicine focused on identifying treatments that promote renal healing after AKI to prevent CKD and remote organ failure.

摘要 多器官功能障碍综合征(MODS)在危重病患者中具有显著的发病率和死亡率。 急性肾损伤(AKI)是MODS最常见的组成部分之一。未解决的AKI导致持续 肾脏损伤、纤维化和随后的慢性肾脏疾病(CKD)。然而，指导肾脏的机制 恢复和防止AKI向CKD过渡的机制还知之甚少。一种新颖而有希望的治疗方法 方法是调节聚集在损伤体内的巨噬细胞和树突状细胞(DC)的功能 肾脏，以防止进一步的损伤和纤维化。白介素1α/β是典型的促炎因子 巨噬细胞和DC与其细胞表面受体IL-1R1结合后激活的细胞因子。IL-1R1 活化在肾脏愈合中起着重要作用。而IL-1R1‘S对巨噬细胞和DC的特异性作用 AKI之后的功能以及AKI到CKD的过渡过程中的作用尚不清楚。根据我们的初步研究， 我们的中心假设是在急性肾损伤后，巨噬细胞IL-1R1的激活触发了肿瘤坏死因子α依赖的肾脏 细胞坏死性下垂，引导依赖IL-1R1的DC向淋巴结迁移以激活促炎作用 T细胞驱动AKI向CKD的转变。我们将对我们的中心假设进行如下测试： 特异性目标1：确定巨噬细胞IL-1R1激活对肿瘤坏死因子介导的肾细胞的影响 缺血性AKI后的坏死性下垂。巨噬细胞特异性缺失IL-1R1(IL-1R1MKO)和 对照组(IL-1R1MWT)将接受缺血/再灌注(I/R)和脓毒症AKI，并观察急性 肾脏损害将被量化。肾细胞坏死性下垂将在受损的肾脏和 巨噬细胞-肾小管细胞(RTC)共培养。我们将采用细胞表面表型和单细胞 从受损肾脏中分离的白细胞的RNA-SEQ精确表型髓系细胞以确定如何 IL-1R1激活调节巨噬细胞极化和肾脏损伤。 特异性目标2：阐明IL-1R1介导的树突状细胞迁移和T细胞活化的作用 在AKI向CKD过渡期间。DC中IL-1R1缺失的小鼠(IL-1R1DCKO)和对照组(IL-1R1 DCWT)将受到I/R诱导的AKI，在多个时间点，肾脏损害的严重程度和 纤维化将被比较。我们将检测IL-1R1介导的DC亚群在肾淋巴中的聚集 I/R后的结节及其对效应器淋巴细胞的激活。我们将使用新的纸巾 流式细胞术分析纤维化区DC和T细胞的空间分布。激活的DC和T细胞将 与RTC共培养，以确定它们对促纤维化基因表达程序的影响。 建议的研究将对促进未来有针对性的发展产生重大的积极影响 对危及生命的AKI患者的治疗。再加上我的培训计划，这些研究将为 作为重症监护医学领域的一名独立研究员，我的发展基础主要是确定 促进AKI后肾脏愈合的治疗，以预防CKD和远端器官衰竭。