Neutrophils in Recovery after ICH

ICH后中性粒细胞的恢复

• 批准号: 9816107
• 负责人: Jaroslaw Aronowski
• 金额: $ 7.37万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-15 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9816107
• 关键词: Affect; Animals; Anti-inflammatory; Apoptosis; Apoptotic; Binding; Biological; Blood; Bone Marrow; Brain; Brain Injuries; CD36 gene; Cells; Cerebral hemisphere hemorrhage; Chemicals; Clinical Trials; Cytolysis; Cytoplasmic Granules; Drug Metabolic Detoxication; Enzymes; Exhibits; Extravasation; Gelatinase B; Goals; Haptoglobins; Hematoma; Heme; Heme Iron; Hemoglobin; Hemolysis; Hemopexin; Hour; Individual; Inflammatory Response; Interleukins; Iron; Ischemic Stroke; LCN2 gene; Lactoferrin; Leukocytes; Mediating; Microglia; Modeling; Modification; Mus; NADPH Oxidase; Necrosis; Neutrophil Infiltration; Neutrophilic Infiltrate; Patients; Phagocytes; Phagocytosis; Phenotype; Pilot Projects; Poison; Primary Cell Cultures; Process; Property; Proteins; Recombinant Interleukins; Recombinants; Recovery; Role; Secondary to; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Source; Therapeutic; Toxic effect; base; brain parenchyma; brain repair; brain tissue; cytotoxic; cytotoxicity; effective therapy; functional loss; immunoregulation; macrophage; microbial; neutrophil; new therapeutic target; novel; novel therapeutics; oxidative damage; scavenger receptor; surface coating; treatment strategy

Shortly after the onset of intracerebral hemorrhage (ICH) and then for several days, masses of polymorphonuclear neutrophils (PMNs) enter the ICH-affected brain. After releasing various cytotoxic moieties, the PMNs undergo rapid apoptosis and then are removed by brain microglia/macrophages (MΦ). Without fast clearance by MΦ, apoptotic PMNs (ANs) proceed to secondary necrosis and spillage of wealth of uniquely toxic moieties, causing harm to brain and augment brain injury caused by hematoma. However, given the biological properties of individual moieties released by PMNs, it is likely that PMNs can also add to brain repair. The PMN's chemical composition is set at the precursor stage of PMNs (pPMN) in bone marrow (BM)(BM-pPMN). The mature PMNs retain the pre-established phenotype and act as shuttles, delivering the content (packed in granules) to the sites of PMNs' infiltration (here ICH-injured brain). Here, we propose to study a novel pathway where the signal, specifically IL-27, modifies BM-pPMN phenotype, so that the mature PMNs upon entering the ICH-affected brain would cause less damage, or even provide benefit, including by promoting phagocytosis-mediated cleanup of hematoma and ANs. Lactoferrin (LTF) is a pleiotropic iron-binding/immunoregulatory protein that is synthetized/distributed by PMNs. Our pilot studies show that ICH-induced IL-27 is a signal for BM-pPMN to express more LTF. The LTF- enriched PMNs upon entering the hematoma could release more LTF. This LTF is capable of: (1) stimulating ANs engulfment by MΦ, (2) upregulating MΦ' CD91, a scavenger receptor for engulfment of LTF-coated ANs and other hemolysis products, (3) sequestrating the free iron, and (4) polarizing MΦ to beneficial (M2-like) phenotype. In addition to LTF, IL-27 promotes expression of hemoglobin neutralizing protein haptoglobin, “help-me-signal” molecule lipocalin-2, and suppresses the expression of MMP-9 and NADPH-oxidase by PMNs. Ultimately, treating animals with recombinant IL-27 (rIL-27) or with recombinant LTF (rLTF), robustly, with a 24h therapeutic window (for rLTF), reduces brain damage and functional loss caused by ICH. Our hypothesis is that after ICH, lL-27 polarizes PMNs to a beneficial type, which upon reaching ICH- injured brain is less cytotoxic and even enhances (in part through LTF) MΦ-mediated cleanup (in part through upregulating CD91/CD36 scavenger receptor) and lessens secondary brain damage. Specific aims are: (1) characterize the IL-27-induced PMN phenotype within BM and blood; (2) with primary cell culture, to define the beneficial role of the PMN phenotype and LTF in clearance/detoxification of ANs/heme/iron by the PMNs/LTF- modified MΦ; and (3) to assess the therapeutic role for rLTF and to define the role of CD91in LTF-assisted cleanup, in mouse ICH model. Our goal is to develop new therapeutic targets for ICH, based on the ability of IL-27-modified PMNs to release more LTF to promote MΦ-mediated cleanup of hematoma. This approach should not only limit PMNs-mediated CNS damage, but also allow PMNs to act in a beneficial fashion.

脑出血（ICH）发生后不久，随后几天内，大量 多形核中性粒细胞 (PMN) 进入受 ICH 影响的大脑。释放各种细胞毒性部分后， PMN 经历快速凋亡，然后被脑小胶质细胞/巨噬细胞 (MΦ) 清除。不带快 通过 MΦ 清除，凋亡的 PMN（AN）继续继发性坏死和独特的财富溢出 有毒部分，对大脑造成伤害并加重血肿引起的脑损伤。 然而，考虑到 PMN 释放的各个部分的生物学特性，PMN 很可能 还可以促进大脑修复。 PMN 的化学成分是在 PMN 的前体阶段 (pPMN) 设定的 骨髓（BM）（BM-pPMN）。成熟的 PMN 保留了预先建立的表型并充当穿梭机， 将内容物（包装在颗粒中）输送到 PMN 渗透的部位（此处为 ICH 损伤的大脑）。 在这里，我们建议研究一种新的途径，其中信号，特别是 IL-27，修饰 BM-pPMN 表型，因此成熟的 PMN 在进入 ICH 影响的大脑时会造成较小的损害，甚至 提供益处，包括促进吞噬作用介导的血肿和 AN 清除。 乳铁蛋白 (LTF) 是一种多效性铁结合/免疫调节蛋白，由以下物质合成/分布 PMN。我们的初步研究表明，ICH 诱导的 IL-27 是 BM-pPMN 表达更多 LTF 的信号。 LTF- 进入血肿后富集的 PMN 可以释放更多的 LTF。该 LTF 能够： (1) 刺激 MΦ 吞噬 AN，(2) 上调 MΦ' CD91，一种吞噬 LTF 包被的 AN 的清道夫受体 和其他溶血产物，(3) 隔离游离铁，以及 (4) 将 MΦ 极化为有益的（类 M2） 表型。除了 LTF 之外，IL-27 还可促进血红蛋白中和蛋白触珠蛋白的表达， “help-me-signal”分子 lipocalin-2，并通过抑制 MMP-9 和 NADPH 氧化酶的表达 PMN。最终，用重组 IL-27 (rIL-27) 或重组 LTF (rLTF) 强有力地治疗动物， 具有 24 小时治疗窗口（对于 rLTF），可减少 ICH 引起的脑损伤和功能丧失。 我们的假设是，在 ICH 之后，IL-27 将 PMN 极化为有益类型，在达到 ICH 后，IL-27 将 PMN 极化为有益类型。 受伤的大脑细胞毒性较小，甚至增强（部分通过 LTF）MΦ 介导的清除（部分通过 上调 CD91/CD36 清道夫受体）并减轻继发性脑损伤。具体目标是：（1） 表征 BM 和血液中 IL-27 诱导的 PMN 表型； (2) 通过原代细胞培养，确定 PMN 表型和 LTF 在 PMN/LTF 对 AN/血红素/铁的清除/解毒中的有益作用- 修改MΦ； (3) 评估 rLTF 的治疗作用并确定 CD91 在 LTF 辅助中的作用 小鼠 ICH 模型中的清理。我们的目标是基于 ICH 的能力开发新的治疗靶点 IL-27 修饰的 PMN 释放更多的 LTF，以促进 MΦ 介导的血肿清除。这种做法 不仅应该限制中性粒细胞介导的中枢神经系统损伤，而且还应该允许中性粒细胞以有益的方式发挥作用。