Cardiac Macrophages as Disease Drivers in Chronic Ischemic Heart Failure

心脏巨噬细胞作为慢性缺血性心力衰竭的疾病驱动因素

基本信息

批准号：
10613345
负责人：
Sumanth D Prabhu
金额：
$ 52.12万
依托单位：
WASHINGTON UNIVERSITY
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：
2021-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10613345
关键词：
Adoptive Cell Transfers Adoptive Transfer Antibodies Antisense Oligonucleotides Attenuated Behavior Biological Assay Biological Response Modifiers Bone Marrow C57BL/6 Mouse Cardiac Cell Proliferation Cell Separation Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Characteristics Chronic Classification Clinical Congestive Heart Failure Coronary Data Development Disease Disease Progression Disease model Exhibits Female Fibrosis Flow Cytometry Functional disorder Genes Genetic Genetic Transcription Heart Heart Injuries Heart failure Human Immune Immunohistochemistry Inflammation Inflammatory Injections Interleukin 4 Receptor Interleukin-10 Interleukin-13 Interleukin-4 Ischemia Knockout Mice Late Effects Left Ventricular Remodeling Ligation Link Macrophage Measures Mediator Modeling Mus Myelogenous Myocardial Infarction Myocardial Ischemia Myocarditis Myocardium Myofibroblast PTPRC gene Pathogenesis Pathologic Pathway interactions Phagocytes Phagocytosis Population Prognosis Progressive Disease Proliferating Proteins Role Signal Pathway Signal Transduction Sorting Spatial Distribution Tamoxifen Testing Therapeutic Tissue Expansion Tissues beta-Chemokines cell type chemokine chemokine receptor cytokine gamma-Chemokines heart cell immune activation immunomodulatory strategy immunoregulation in vivo innovation insight ischemic cardiomyopathy kinase inhibitor male monocyte neutralizing antibody novel novel strategies recruit repaired self renewing cell self-renewal single-cell RNA sequencing small molecule source localization targeted treatment therapeutic target tissue injury transcriptomics virtual

项目摘要

Macrophage expansion in the failing heart induces tissue injury and is thought to contribute to the progression of heart failure (HF). The main contributors to macrophage expansion in the failing heart are cells that self-renew and proliferate independent of the blood monocyte pool (and hence are C-C chemokine receptor 2 [CCR2]–). However, the role of such locally-sourced macrophages in the pathogenesis of chronic left ventricular (LV) remodeling is poorly understood. The normal heart harbors macrophages expressing CD206 (Mrc1), along with the gene markers Ym1, Fizz1, and Arg1, that are primarily CCR2–. These CD206+ macrophages can be activated by Th2 cytokines such as interleukin(IL)-4/IL-13. Moreover, secreted FIZZ1 contributes to myofibroblast activation and fibrosis in other disease models. Our pilot data indicate that CD206+ macrophages expressing IL- 4 receptora (IL-4Ra) robustly increase in the failing heart, and suggest that macrophage IL-4/IL-4Ra signaling promotes LV remodeling and fibrosis, in part through FIZZ1. Hence, we hypothesize that cardiac CD206+ macrophages expressing IL-4Ra and Fizz1 are innate immune mediators of adverse LV remodeling in chronic HF, and key targets for therapeutic immunomodulation. Three Aims will test this hypothesis. In Aim 1, in a murine coronary ligation model, using flow cytometry, cell sorting, single cell RNA sequencing (scRNAseq), and immunohistochemistry, we will comprehensively define pathological alterations in cardiac CD206+ macrophages in HF, including IL-4Ra levels and downstream signaling, FIZZ1 expression, and in vivo cell abundance, proliferation, turnover and phagocytic capacity. Using scRNAseq, we will define novel functional subpopulations of CD206+ macrophages at the transcriptional level in both murine and human failing hearts. In Aim 2, we will establish the pathogenetic role of CD206+IL-4Ra+ macrophages in HF using CD45.2 inducible myeloid-specific IL-4Ra knockout mice, deleting myeloid IL-4Ra during chronic HF, and assessing the late effects on LV remodeling, inflammation, and fibrosis. To establish sufficiency of failing heart CD206+IL-4Ra+ macrophages to induce tissue injury, we will adoptively transfer sorted cardiac CD206+ macrophages with intact or deleted IL- 4Ra from HF mice into naïve CD45.1 mice via intramyocardial injection and assess LV remodeling 6 w later. To assess the role of FIZZ1, we will similarly transfer M[IL-4] polarized bone marrow macrophages from wild-type and Fizz1-/- mice. In Aim 3, we will test potentially translatable therapies to antagonize CD206+IL-4Ra+ macrophages and alleviate chronic LV remodeling, including systemic anti-sense oligonucleotides against IL- 4Ra and Fizz1, anti-IL-4 neutralizing antibody, and GW2580, a small molecule cFMS kinase inhibitor. We will measure the effects of these therapies on LV remodeling, cardiac macrophages, fibrosis, and chemokine/ cytokine expression. These studies will further our understanding of the innate immune basis for cardiac inflammation in ischemic HF, provide novel insights into macrophage IL-4-dependent signaling and Fizz1 in the pathogenesis of LV remodeling, and identify new approaches for cell type-specific immunomodulation in HF.

衰竭心脏中巨噬细胞的扩张会引起组织损伤，并被认为有助于病情进展心力衰竭（HF）中巨噬细胞扩张的主要贡献者是自我更新的细胞。并独立于血液单核细胞库进行增殖（因此是 C-C 趋化因子受体 2 [CCR2]–）。然而，这种本地来源的巨噬细胞在慢性左心室（LV）发病机制中的作用人们对正常心脏的重塑知之甚少，巨噬细胞表达 CD206 (Mrc1)。基因标记 Ym1、Fizz1 和 Arg1（主要是 CCR2-）可以被激活。 Th2 细胞因子如白细胞介素 (IL)-4/IL-13 此外，分泌的 FIZZ1 有助于肌成纤维细胞。我们的试验数据表明 CD206+ 巨噬细胞表达 IL- 4 受体 a (IL-4Ra) 在衰竭心脏中急剧增加，表明巨噬细胞 IL-4/IL-4Ra 信号传导部分通过 FIZZ1 促进左心室重塑和纤维化因此，我们与心脏 CD206+ 作斗争。表达 IL-4Ra 和 Fizz1 的巨噬细胞是慢性不良左心室重塑的先天免疫介质 HF 和治疗性免疫调节的关键目标将在目标 1 中在小鼠中验证这一假设。冠状动脉结扎模型，使用流式细胞术、细胞分选、单细胞 RNA 测序 (scRNAseq) 和免疫组织化学，我们将全面定义心脏CD206+巨噬细胞的病理改变 HF 中，包括 IL-4Ra 水平和下游信号传导、FIZZ1 表达和体内细胞丰度，使用 scRNAseq，我们将定义新的功能亚群。在目标 2 中，我们将在小鼠和人类衰竭心脏的转录水平上检测 CD206+ 巨噬细胞。使用 CD45.2 诱导骨髓特异性确定 CD206+IL-4Ra+ 巨噬细胞在心力衰竭中的致病作用 IL-4Ra 基因敲除小鼠，在慢性心力衰竭期间删除骨髓 IL-4Ra，并评估对 LV 的后期影响重塑、炎症和纤维化确定衰竭心脏 CD206+IL-4Ra+ 巨噬细胞的充足性。诱导组织损伤，我们将过继转移具有完整或删除的 IL- 的分类心脏 CD206+ 巨噬细胞通过心肌内注射将 4Ra 从 HF 小鼠转移到幼稚 CD45.1 小鼠中，并在 6 周后评估 LV 重塑。为了评估 FIZZ1 的作用，我们将类似地从野生型转移 M[IL-4] 极化骨髓巨噬细胞在目标 3 中，我们将测试拮抗 CD206+IL-4Ra+ 的潜在可转化疗法。巨噬细胞并缓解慢性左室重塑，包括针对 IL- 的全身反义寡核苷酸 4Ra 和 Fizz1（抗 IL-4 中和抗体）和 GW2580（小分子 cFMS 激酶抑制剂）。测量这些疗法对左室重塑、心脏巨噬细胞、纤维化和趋化因子的影响/ 这些研究将进一步了解心脏的先天免疫基础。缺血性心力衰竭中的炎症，为巨噬细胞 IL-4 依赖性信号传导和 Fizz1 的新见解提供了新的见解左室重塑的发病机制，并确定心力衰竭细胞类型特异性免疫调节的新方法。