Complement as a modulator of immunosuppression and progression in Polycystic Kidney Disease

补体作为多囊肾病免疫抑制和进展的调节剂

• 批准号: 10282996
• 负责人: Katharina Hopp
• 金额: $ 11.66万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10282996
• 关键词: Adaptive Immune System; Anaphylatoxins; Automobile Driving; Autosomal Dominant Polycystic Kidney; Binding; C3AR1 gene; C5a anaphylatoxin receptor; CD8-Positive T-Lymphocytes; Cancer Model; Cell Separation; Cells; Cleaved cell; Clinical Trials; Complement; Complement 3a; Complement 5a; Complement Activation; Complement Inactivators; Complement Membrane Attack Complex; Cyst; Cyst Fluid; Cystic kidney; DNA Sequence Alteration; Data; Disease; Disease model; End stage renal failure; Epithelial; Epithelial Cells; FDA approved; Flow Cytometry; Future; Genetic; Goals; Grant; Growth; Image; Immune; Immune Evasion; Immune checkpoint inhibitor; Immunity; Immunosuppression; Immunotherapy; Impairment; Inherited; Innate Immune System; Investigation; K-Series Research Career Programs; Kidney; Kidney Diseases; Lead; Malignant Neoplasms; Mediating; Metabolic; Methods; Modeling; Molecular; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Outcome; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Polycystic Kidney Diseases; Pre-Clinical Model; Production; Proteins; Publications; Publishing; Quality of life; Receptor Inhibition; Regimen; Regulatory T-Lymphocyte; Reporting; Research; Research Personnel; Role; Series; Severities; Severity of illness; Shapes; Signal Transduction; T-Lymphocyte; Testing; Therapeutic; Time; Translating; Tumor Burden; Tumor Escape; Tumor-associated macrophages; Up-Regulation; Work; adaptive immunity; cancer immunotherapy; cell type; clinically relevant; complement pathway; cytokine; fluorophore; immune checkpoint; inhibitor/antagonist; innovation; macrophage; mouse model; new therapeutic target; overexpression; pre-clinical; pre-clinical research; receptor; therapeutic evaluation; therapeutic target; translational research program; tumor progression; tumor-immune system interactions

PROJECT SUMMARY Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common, monoallelic nephropathy worldwide, characterized by continuous renal cysts growth leading to end stage kidney disease. A single FDA approved therapy is available, but it merely slows cyst growth and impacts quality of life, highlighting the urgent need for new treatment options. ADPKD presents with high phenotypic variability, suggesting that mechanisms beyond the genetic mutation to either PKD1 or PKD2 influence disease severity. Recent data have implicated immune cells as modulators of disease severity. Multiple publications highlight that M2-like renal macrophages promote cystic progression. Conversely, data from my K01, using the slowly progressive, orthologous Pkd1 p.R3277C (Pkd1RC/RC) ADPKD model, show that CD8+ T cells can inhibit renal cyst growth. We further found that the renal cystic microenvironment displays multiple features of immunosuppression such as increased numbers of CD4+ regulatory T cells, metabolic reprogramming, and engagement of immune checkpoints. Indeed, immune checkpoint inhibitors, which lead to reactivation of CD8+ T cells, alleviate cystic disease in the Pkd1RC/RC model. What remains unknown are the mechanisms that drive immunosuppression, which may present important/novel therapeutic targets. In cancer, a disease that parallels PKD at the cellular and molecular level, activation of the complement pathway has been shown to promote an immunosuppressive microenvironment. The complement cascade is a central part of the innate immune system that regulates adaptive immunity. Complement has been shown to be upregulated in PKD cells, patient kidneys/cyst fluid, and murine models, including the Pkd1RC/RC mouse as shown by our preliminary data. Further, genetic loss or non-selective inhibition of the complement protein C3 slows cyst growth in murine PKD models. We hypothesize that complement signaling drives PKD, in part, by creating an immunosuppressive microenvironment which impairs the adaptive immune system to halt cyst growth. This project has two aims. In Aim 1, we will utilize fluorescent-activated cell sorting, flow cytometry, and multispectral immunofluorescent imaging to determine which cells are key producers and responders of complement signaling in the Pkd1RC/RC kidney. We will further correlate these findings to disease severity. In Aim 2, we will test if targeted complement inhibition alleviates cystic disease. This will be the first of such studies using a clinically relevant, specific complement inhibitor in a model orthologous to ADPKD, the Pkd1RC/RC mouse. We will further utilize sophisticated 64-fluorophore capable flow cytometry methods to investigate how complement inhibition alters the cystic immune microenvironment with specific focus on immunosuppressive features. These investigations directly expand upon my NIDDK K01 work and will provide key preliminary data for a competitive R01 application. Obtaining this R03 will further facilitate my transition into an independent investigator with an innovative, preclinical, and translational research program that focuses on the role of the cystic microenvironment as modulator of and therapeutic target for PKD.

项目摘要 常染色体显性多囊性肾脏疾病（ADPKD）是最常见的单相肾病 在全球范围内，其特征是连续的肾囊肿生长，导致末期肾脏疾病。一个FDA 有批准的疗法可用，但它只是减慢了囊肿的生长并影响生活质量，突出了紧急的 需要新的治疗选择。 ADPKD呈现高表型变异性，表明机制 除了PKD1或PKD2的遗传突变外，会影响疾病的严重程度。最近的数据牵涉 免疫细胞作为疾病严重程度的调节剂。多个出版物强调了类似M2的肾脏巨噬细胞 促进囊性进展。相反，我的K01的数据，使用缓慢进行的直系同源PKD1 P.R3277C（PKD1RC/RC）ADPKD模型，表明CD8+ T细胞可以抑制肾脏囊肿的生长。我们进一步发现 肾囊性微环境显示了免疫抑制的多个特征，例如增加数量 CD4+调节T细胞，代谢重编程和免疫检查点的参与度。确实，免疫 检查点抑制剂，导致CD8+ T细胞重新活化，可减轻PKD1RC/RC模型中的囊性疾病。 尚不清楚的是驱动免疫抑制的机制，这些机制可能呈现重要/新颖 治疗靶标。在癌症中，一种在细胞和分子水平上与PKD相似的疾病，激活 补体途径已显示可促进免疫抑制微环境。补充 级联是调节适应性免疫的先天免疫系统的中心部分。补充已经是 显示在PKD细胞，患者肾脏/囊肿液和鼠模型中上调，包括PKD1RC/RC 鼠标如我们的初步数据所示。此外，遗传丧失或对补体的非选择性抑制 蛋白C3在鼠PKD模型中减慢了囊肿的生长。我们假设补体信号传导驱动PKD， 一部分，通过创建一种免疫抑制微环境，损害适应性免疫系统以停止 囊肿生长。该项目有两个目标。在AIM 1中，我们将利用荧光激活的细胞分选，流式细胞仪， 和多光谱免疫荧光成像，以确定哪些细胞是关键生产者和反应者 PKD1RC/RC肾脏中的补体信号传导。我们将进一步将这些发现与疾病的严重程度相关联。目标 2，我们将测试是否有针对性的补体抑制减轻囊性疾病。这将是此类研究的第一项 在与PKD1RC/RC小鼠的ADPKD模型中，使用临床相关的特定补体抑制剂。 我们将进一步利用复杂的64-氟化合物流式细胞术方法来研究如何 补体抑制作用改变了囊性免疫微环境，专门针对免疫抑制 特征。这些调查直接扩展了我的NIDDK K01工作，并将提供关键的初步数据 用于竞争性R01应用程序。获得此R03将进一步促进我向独立的过渡 具有创新，临床前和转化研究计划的研究人员，重点是 囊性微环境作为PKD的调节剂和治疗靶标。