Proangiogenic M2-type macrophages and choroidal neovascularization

促血管生成 M2 型巨噬细胞和脉络膜新生血管

基本信息

批准号：
10681491
负责人：
Alexander Georg Marneros
金额：
$ 42.38万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681491
关键词：
ABL1 gene Ablation Address Angiogenesis Inhibition Angiogenesis Inhibitors Antigen-Antibody Complex Aqueous Humor Area Attenuated Biological Assay Blood Vessels Cells Chemicals Choroidal Neovascularization Data Data Analyses Disease Drug Screening Endothelium Event Exudative age-related macular degeneration Eye Genetic Glucocorticoids Heat-Shock Proteins 90 Histone Deacetylase Histone Deacetylase Inhibitor Impairment In Vitro Infiltration Inflammation Inflammatory Injections Interleukin-10 Interleukin-13 Interleukin-4 Interleukin-6 Lasers Lesion MAP2K1 gene MEKs Macrophage Mediating Membrane Modeling Molecular Molecular Target Mus Pathogenesis Patients Pharmaceutical Preparations Phase Phosphotransferases Population Process Proteomics Resources Role Signal Transduction Site Testing Therapeutic Time VEGFA gene angiogenesis cellular targeting clinically relevant cytokine experimental study genetic approach in vivo inhibitor innovation macrophage-derived chemokine mouse model neovascularization novel novel strategies novel therapeutic intervention novel therapeutics pharmacologic phosphoproteomics prevent small molecule therapeutic target

项目摘要

SUMMARY Macrophages infiltrate the site of inflammation and polarize to either antiangiogenic M1-type macrophages or proangiogenic M2-type macrophages, depending on the presence of various cytokines and other external factors. Proangiogenic M2-type macrophages are major contributors to inflammatory angiogenesis in many common diseases, in part by secreting proangiogenic factors, such as VEGF-A or IL-1b. Notably, M2-type macrophages accumulate in areas of choroidal neovascularization (CNV) in patients with neovascular AMD. The increase in M2-type macrophage-derived cytokines in eyes of patients with neovascular AMD has implicated this cell population as a major driver of CNV pathogenesis. This hypothesis is further supported by the observation of an accumulation of M2-type macrophages in laser-induced CNV or in a genetic VEGF-A-induced mouse model of neovascular AMD. Ablation of macrophages in laser-induced CNV, which are predominantly M2-type macrophages, blocked CNV, whereas administration of M2-type macrophages in the eye promoted CNV. Based on these findings we propose that pharmacologic therapies that prevent M2-type polarization of macrophages can serve as a novel approach to potently inhibit CNV progression in patients with neovascular AMD. The limited understanding of the signaling mechanisms that are regulating M1- versus M2-type polarization of macrophages has hindered the identification of molecular targets and pharmacologic inhibitors to selectively block M2-type macrophage polarization. To address this unmet need, we have performed global quantitative time-course proteomics and phosphoproteomics and identified kinase activation events that are associated with M1- versus M2-type macrophage polarization. Furthermore, we identified in chemical screens pharmacologic inhibitors of M2-type polarization that selectively block M2- but not M1-type polarization. Thus, the combination of these two approaches provides us now with a unique resource to establish novel therapies that selectively block proangiogenic M2- but not antiangiogenic M1-type macrophages in CNV. In proof-of-principle experiments we can show that two of the identified inhibitors, the MEK inhibitor trametinib and the HDAC inhibitor panobinostat, blocked M2-type macrophage polarization in CNV lesions and potently inhibited CNV lesion formation in laser- induced CNV experiments. Our proposed experiments will test identified kinase and non-kinase inhibitors in a systematic manner for their ability to selectively block M2-type polarization in detailed in vitro experiments as well as in three well-established mouse models of neovascular AMD. Our proposal is based on extensive preliminary data that provide a strong scientific premise. The proposed experiments have high rigor and important clinical relevance and will likely lead to novel therapeutic approaches for neovascular AMD.

概括巨噬细胞浸润炎症部位并极化为抗血管生成 M1 型巨噬细胞或促血管生成 M2 型巨噬细胞，取决于各种细胞因子和其他外部因素的存在因素。促血管生成 M2 型巨噬细胞是许多炎症血管生成的主要贡献者常见疾病的发生部分是通过分泌促血管生成因子（例如 VEGF-A 或 IL-1b）来实现的。值得注意的是，M2型新生血管性 AMD 患者的脉络膜新生血管 (CNV) 区域会积聚巨噬细胞。这新生血管性 AMD 患者眼中 M2 型巨噬细胞衍生细胞因子的增加暗示了这一点细胞群是 CNV 发病机制的主要驱动力。这一假设得到了观察的进一步支持激光诱导的 CNV 或遗传 VEGF-A 诱导的小鼠中 M2 型巨噬细胞的积累新生血管AMD模型。激光诱导 CNV 中巨噬细胞的消融，主要是 M2 型巨噬细胞阻断 CNV，而在眼睛中施用 M2 型巨噬细胞则促进 CNV。基于根据这些发现，我们建议采用药物疗法来预防巨噬细胞的 M2 型极化可以作为一种有效抑制新生血管性 AMD 患者 CNV 进展的新方法。有限的了解调节巨噬细胞 M1 型与 M2 型极化的信号传导机制阻碍了选择性阻断 M2 型的分子靶点和药物抑制剂的鉴定巨噬细胞极化。为了解决这一未满足的需求，我们进行了全球定量时间进程蛋白质组学和磷酸化蛋白质组学，并鉴定了与 M1- 相关的激酶激活事件 M2型巨噬细胞极化。此外，我们在化学筛选中发现了药物抑制剂 M2 型极化，选择性阻止 M2 型极化，但不阻止 M1 型极化。因此，这两者的结合方法现在为我们提供了独特的资源来建立选择性阻断的新疗法 CNV 中促血管生成 M2 型巨噬细胞，但不具有抗血管生成 M1 型巨噬细胞。在原理验证实验中我们可以显示两种已确定的抑制剂，MEK 抑制剂曲美替尼和 HDAC 抑制剂帕比司他，阻断 CNV 病变中的 M2 型巨噬细胞极化，并有效抑制激光中 CNV 病变的形成诱导CNV实验。我们提出的实验将测试已识别的激酶和非激酶抑制剂在详细的体外实验中系统地评价其选择性阻断 M2 型极化的能力：以及三种成熟的新生血管 AMD 小鼠模型。我们的建议是基于广泛的初步数据提供了强有力的科学前提。所提出的实验具有很高的严谨性和重要的临床意义，可能会导致新生血管性 AMD 的新治疗方法。