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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：
10228245
负责人：
Sumanth D Prabhu
金额：
$ 49.61万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
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 2021-09-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10228245
关键词：
Acute 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 Disease Disease Progression Disease model Exhibits Female Fibrosis Flow Cytometry Functional disorder Genes Genetic Transcription Heart Heart Injuries Heart failure Human Immune Immunohistochemistry Inflammation Inflammatory Injections Interleukin Receptor Interleukin-10 Interleukin-13 Interleukin-4 Interleukins Knockout Mice Late Effects Left Ventricular Remodeling Ligation Link Measures Mediator of activation protein 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 Source Spatial Distribution Tamoxifen Testing Therapeutic Tissue Expansion Tissues beta-Chemokines cardiogenesis cell type chemokine chemokine receptor cytokine gamma-Chemokines heart cell immune activation immunomodulatory strategy immunoregulation in vivo innovation insight ischemic cardiomyopathy kinase inhibitor macrophage male monocyte neutralizing antibody novel novel strategies recruit repaired self renewing cell self-renewal single-cell RNA sequencing small molecule 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.

衰竭心脏中的巨噬细胞扩增诱导组织损伤，并被认为有助于进展心力衰竭（HF）。衰竭心脏中巨噬细胞扩张的主要贡献者是自我更新的细胞并且不依赖于血液单核细胞池增殖（因此是C-C趋化因子受体2 [CCR 2]-）。然而，这种局部来源的巨噬细胞在慢性左心室（LV）重塑是知之甚少。正常心脏含有表达CD 206（Mrc 1）的巨噬细胞，沿着基因标记Ym 1、Fizz 1和Arg 1，主要是CCR 2-。这些CD 206+巨噬细胞可以被激活， Th 2细胞因子如白细胞介素（IL）-4/IL-13。此外，分泌的FIZZ 1有助于肌成纤维细胞活化和纤维化。我们的初步数据表明，表达IL-10的CD 206+巨噬细胞在衰竭的心脏中，巨噬细胞IL-4/IL-4 Ra信号转导显著增加，促进LV重塑和纤维化，部分通过FIZZ 1。因此，我们假设心脏CD 206 + 表达IL-4 Ra和Fizz 1的巨噬细胞是慢性心肌梗死患者左室重构的先天性免疫介质 HF和治疗性免疫调节的关键靶点。三个目标将检验这一假设。在Aim 1中，在小鼠中，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），免疫组化，我们将全面定义心脏CD 206+巨噬细胞的病理变化，在HF中，包括IL-4 Ra水平和下游信号传导、FIZZ 1表达和体内细胞丰度，增殖、周转和吞噬能力。使用scRNAseq，我们将定义新的功能亚群， CD 206+巨噬细胞在小鼠和人类衰竭心脏中的转录水平。在目标2中，我们将使用CD45.2诱导的骨髓特异性免疫荧光技术， IL-4 Ra敲除小鼠，在慢性HF期间删除髓样IL-4 Ra，并评估对LV的晚期影响重塑炎症和纤维化确定衰竭心脏CD 206 +IL-4 Ra+巨噬细胞的充足性，诱导组织损伤，我们将过继性转移分选的心脏CD 206+巨噬细胞与完整或缺失的IL- 4 Ra通过心肌内注射到未经处理的CD 45.1小鼠中，并在6周后评估LV重构。到为了评估FIZZ 1的作用，我们将类似地将M[IL-4]极化的骨髓巨噬细胞从野生型和Fizz 1-/-小鼠。在目标3中，我们将测试拮抗CD 206 +IL-4 Ra+的潜在可转化疗法。巨噬细胞和减轻慢性LV重塑，包括针对IL-12的系统性反义寡核苷酸。 4 Ra和Fizz 1，抗IL-4中和抗体，和GW 2580，一种小分子cFMS激酶抑制剂。我们将测量这些疗法对LV重塑、心脏巨噬细胞、纤维化和趋化因子的影响。细胞因子表达这些研究将进一步加深我们对心脏病先天免疫基础的理解。缺血性HF炎症，提供了新的见解巨噬细胞IL-4依赖性信号和Fizz 1在 LV重构的发病机制，并确定HF中细胞类型特异性免疫调节的新方法。