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: 10605321
• 负责人: Jamie R Privratsky
• 金额: $ 15.81万
• 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/10605321
• 关键词: Acceleration; Acute; Acute Renal Failure with Renal Papillary Necrosis; Admission activity; 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 Activation; Interleukin-1 Receptors; Interleukins; Ischemia; Kidney; Knowledge; Leukocytes; Life; Lymphocyte; Lymphocyte Activation; Macrophage; Measures; Mediating; Mediator; Medicine; Methods; Mission; Modeling; Morbidity - disease rate; Multiple Organ Failure; Mus; Myeloid Cells; Nature; Operative Surgical Procedures; Organ; 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; Sorting; 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; body system; cell injury; cell motility; cytokine; disability; draining lymph node; experience; healing; immunomodulatory therapies; immunoregulation; improved; kidney cell; kidney fibrosis; lymph nodes; 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.

抽象的 多器官功能障碍综合征（MODS）导致危重患者显着的发病率和死亡率。 急性肾损伤 (AKI) 是 MODS 最常见的组成部分之一。未解决的 AKI 会导致持续的 肾损伤、纤维化和随后的慢性肾病（CKD）。然而，指导肾脏的机制 恢复和防止 AKI 向 CKD 转变的了解还很少。一种新颖且有前途的治疗方法 方法是调节在受伤部位积聚的巨噬细胞和树突状细胞 (DC) 的功能 肾脏以防止进一步损伤和纤维化。白细胞介素 (IL)-1α/β 是典型的促炎物质 与细胞表面受体 IL-1R1 结合后激活巨噬细胞和 DC 的细胞因子。白细胞介素1R1 激活在肾脏愈合中发挥着重要作用。然而，IL-1R1 对巨噬细胞和 DC 的特定作用 AKI 后的功能以及 AKI 向 CKD 转变的功能尚不清楚。根据我们的初步研究， 我们的中心假设是，AKI 后，巨噬细胞 IL-1R1 激活会触发 TNFα (TNF) 依赖性肾病 细胞坏死性凋亡，引导 DC 依赖 IL-1R1 迁移至淋巴结以激活促炎细胞 T 细胞驱动 AKI 向 CKD 转变。我们将按如下方式检验我们的中心假设： 具体目标 1：确定巨噬细胞 IL-1R1 激活对 TNF 介导的肾细胞的影响 缺血性 AKI 后坏死性凋亡。巨噬细胞特异性删除 IL-1R1 (IL-1R1 MKO) 的小鼠 对照（IL-1R1 MWT）将经历缺血/再灌注（I/R）和脓毒性 AKI，以及急性的严重程度 肾脏损害将被量化。将在受伤的肾脏和在 巨噬细胞-肾小管细胞（RTC）共培养。我们将采用细胞表面表型分析和单细胞 对受损肾脏的分选白细胞进行 RNA 测序，以精确表型骨髓细胞，以确定如何 IL-1R1 激活调节巨噬细胞极化和肾损伤。 具体目标 2：阐明 IL-1R1 介导的树突状细胞迁移和 T 细胞激活的作用 在 AKI 向 CKD 过渡期间。 DC 中 IL-1R1 缺失的小鼠 (IL-1R1 DCKO) 和对照小鼠 (IL-1R1 DCWT）将遭受 I/R 诱导的 AKI，并且在多个时间点，肾损伤的严重程度和 纤维化程度将进行比较。我们将检查肾淋巴中 IL-1R1 介导的 DC 亚群的积累 I/R 后的淋巴结及其随后的效应淋巴细胞激活。我们将使用新型组织 细胞计数分析纤维化区域内 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。 拟议的研究将通过促进未来有针对性的发展产生重大积极影响 危及生命的 AKI 患者的治疗。这些研究与我的培训计划一起奠定了 我作为重症监护医学独立研究者的发展基础专注于识别 促进 AKI 后肾脏愈合的治疗，以预防 CKD 和远端器官衰竭。