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，然后几天内，大量的 多形核中性粒细胞（PMN）进入ICH影响的大脑。在释放各种细胞毒性部分后， PMNs经历快速凋亡，然后被脑小胶质细胞/巨噬细胞（MΦ）去除。无快速 MΦ清除后，凋亡的中性粒细胞（ANs）发生继发性坏死，并溢出大量独特的 毒性部分，对脑造成伤害，加重血肿引起的脑损伤。 然而，考虑到PMN释放的单个部分的生物学特性， 也能促进大脑修复中性粒细胞的化学组成被设定在中性粒细胞的前体阶段（pPMN）， 骨髓（BM）（BM-pPMN）。成熟的中性粒细胞保留了预先建立的表型并充当穿梭者， 将内容物（以颗粒形式包装）递送到PMN浸润的部位（此处为ICH损伤的脑）。 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 因此，成熟的中性粒细胞在进入ICH影响的大脑后会造成更少的损伤，甚至是更小的损伤。 提供益处，包括通过促进吞噬介导的血肿和AN清除。 乳铁蛋白（LTF）是一种多效性铁结合/免疫调节蛋白，其由哺乳动物合成/分布。 PMNs。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- 在进入血肿时富集的PMNs可以释放更多的LTF。该LTF能够：（1）刺激 （2）上调巨噬细胞吞噬LTF包被的ANs的清道夫受体MΦ'CD 91 和其他溶血产物，（3）螯合游离铁，和（4）极化MΦ至有益（M2样） 表型除了LTF外，IL-27还促进血红蛋白中和蛋白结合珠蛋白的表达， “帮助我信号”分子脂质运载蛋白-2，并通过抑制MMP-9和NADPH-氧化酶的表达， PMNs。最终，用重组IL-27（rIL-27）或重组LTF（rLTF）稳健地治疗动物， 具有24小时治疗窗（对于rLTF），减少由ICH引起的脑损伤和功能丧失。 我们的假设是，在ICH后，IL-27将PMN极化为有益类型，其在达到ICH后- 损伤的脑细胞毒性较低，甚至增强（部分通过LTF）MΦ介导的清除（部分通过LTF）。 上调CD 91/CD 36清道夫受体）并减轻继发性脑损伤。具体目标是：（1） 在BM和血液中表征IL-27诱导的PMN表型;（2）用原代细胞培养，以确定 PMN表型和LTF在PMN/LTF清除/解毒AN/血红素/铁中的有益作用- （3）评估rLTF的治疗作用，并确定CD 91在LTF辅助的 清除，在小鼠ICH模型中。我们的目标是开发新的ICH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。这种方法 这不仅限制了PMN介导的CNS损伤，而且还允许PMN以有益的方式发挥作用。