ILC2s Polarize Microglia to a Reparative, M2-Like Phenotype

ILC2 将小胶质细胞极化为修复性 M2 样表型

• 批准号: 10316989
• 负责人: Alexis Mobley
• 金额: $ 1.65万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2022-04-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316989
• 关键词: Acute; Adoptive Cell Transfers; Affect; Age; Age-associated memory impairment; Aging; Amphiregulin; Anti-Inflammatory Agents; Apoptotic; Biological Assay; Biological Response Modifiers; Blood; Brain; Brain Injuries; Bypass; Cause of Death; Cell Count; Cell Survival; Cells; Communication; Conditioned Culture Media; Cytoprotection; Data; Effector Cell; Ensure; Environment; FDA approved; Female; Flow Cytometry; Gatekeeping; Gene Expression; Genes; Glucose; Growth Factor; Harvest; Health; Homeostasis; Immune; Immune response; Immune system; Immunologics; Immunosuppression; In Vitro; Inflammaging; Inflammation; Inflammatory; Inflammatory Response; Injections; Injury; Interleukin-10; Interleukin-13; Interleukin-4; Interleukin-5; Interleukins; Investigation; Ischemia; Kidney; Latex Bead; Literature; Lymphoid Cell; Mediating; Mediator of activation protein; Microglia; Morphology; Mus; Myeloid Cells; Nerve Degeneration; Neuraxis; Neurologic; Neuromedin U; Neurons; Neuropeptides; Oxygen; Pathway Analysis; Patients; Peptide Hydrolases; Peripheral; Persons; Phagocytes; Phagocytosis; Phenotype; Process; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Regulation; Reperfusion Injury; Reperfusion Therapy; Risk Factors; Role; Signal Transduction; Source; Stress; Stroke; Structure; Substance P; System; T-Lymphocyte; TNF gene; Therapeutic; Time; Tissues; Transgenic Mice; Transplantation; United States; Up-Regulation; Vascular Endothelial Growth Factors; Vasoactive Intestinal Peptide; Western Blotting; Wild Type Mouse; aged; aging brain; arginase; brain cell; brain repair; cell type; cytokine; deprivation; effective therapy; experimental study; falls; healing; healthy aging; heme oxygenase-1; improved; in vitro Model; inflammatory milieu; injured; insight; ischemic injury; macrophage; male; mouse model; nervous system disorder; neuroinflammation; neurovascular; novel; pathogen; peptide P; polarized cell; post stroke; protein expression; pup; repaired; reparative process; response; sex; stroke model; stroke outcome; stroke recovery; stroke therapy; targeted treatment; transcriptome; transcriptome sequencing

Project Summary: Stroke is the fifth leading cause of death in the United States, predominantly affecting aged people. FDA approved treatments are proving inadequate as many patients are ineligible for reperfusion therapies and they fail to treat secondary brain injury or help in brain repair. One of the most promising therapies for stroke is targeting the peripheral and central nervous system immune responses that have modulatory roles during all stages of stroke pathobiology. Aging affects immunologic responses by a global suppression of the immune system, including dysregulation of cytokine mediators, leading to increased inflammation throughout all systems, termed inflammaging. However, understanding mechanisms of healthy aging can bypass this effect. Two candidate immune cells (1) microglia, brain-resident macrophages, and (2) innate lymphoid cells, a newly discovered innate immune cell, may control pre- and post-stroke responses in the aged brain. Microglia immunosurveil the brain during homeostasis and are quickly primed once injury is detected. Unfortunately, they are affected by inflammaging causing them to be poor at anti-inflammatory responses, especially after brain injury. Group 2 innate lymphoid cells (ILC2s) are a prime candidate in regulating microglial responses as they produce anti-inflammatory cytokines (IL-4, IL-5, and IL-13) during times of injury. These cytokines modulate microglia to a reparative phenotype by increasing their trophic and phagocytic capacity through upregulation of healing proteins like heme oxygenase 1, arginase 1, and vascular endothelial growth factor alpha. The communication between macrophages and ILC2s have been investigated peripherally in kidney ischemic- reperfusion injury, but very little brain and in aging. Preliminary data show that with age, ILC2s increase in the brain of male and female mice compared to other ILC types. Moreover, males have a multi-fold global increase of total ILC and ILC2 counts putting their total ILC and ILC2 counts greater than females. Also, male ILC2s produce more cytokines and have a greater anti-inflammatory capacity in aging than females. Lastly, young ILC2 soluble factors increase Nrf2 and Nrf2-regulated genes in microglia in vitro. This would suggest that aged ILC2s could be beneficial in microglia polarization during inflammatory responses. Therefore, I hypothesize that ILC2s are the brain’s gatekeepers of microglia polarization to a reparative, M2-like phenotype under: (1) homeostatic conditions, (2) ischemic insult, and (3) with aging. In Aim 1, I will investigate the transcriptome of ILC2s and ILC2 soluble factors on microglia polarization. In Aim 2, I will identify the response of ILC2s to oxygen-glucose deprivation, an in vitro model of ischemic injury, and how their soluble factors polarize primed microglia. Lastly, in Aim 3, I will perform a “proof-of-concept” experiment to inquire the role of ex-vivo stimulated ILC2s on aged microglia via adoptive cell transfer using intracortical injections of GFP+ ILC2s. Taken together, I will have defined the capacity of ILC2 soluble factors to polarize microglia to a reparative phenotype. This will provide the rationale to further explore ILC2 as a possible therapeutic platform in stroke and other neurologic disorders.

项目摘要：中风是美国第五大主要死因，主要影响老年人 人民。FDA批准的治疗被证明是不够的，因为许多患者不符合再灌注条件 治疗方法既不能治疗继发性脑损伤，也不能帮助大脑修复。最有希望的疗法之一 因为中风的目标是具有调节作用的外周和中枢神经系统的免疫反应 在中风病理生物学的所有阶段。衰老通过对免疫反应的整体抑制来影响免疫反应 免疫系统，包括细胞因子介质的失调，导致全身炎症增加 系统，称为发炎。然而，理解健康衰老的机制可以绕过这种影响。 两种候选免疫细胞(1)小胶质细胞，脑内停留的巨噬细胞，和(2)天然淋巴样细胞，一种新的 发现了先天免疫细胞，可以控制老年大脑中风前后的反应。小胶质细胞 在动态平衡期间，免疫监测大脑，一旦检测到损伤，就会迅速做好准备。不幸的是，他们 受到炎症的影响，导致他们在抗炎反应方面很差，特别是在大脑 受伤。第二组先天淋巴样细胞(ILC2s)是调节小胶质细胞反应的首选候选细胞，因为它们 在受伤时产生抗炎细胞因子(IL-4、IL-5和IL-13)。这些细胞因子调节 通过上调小胶质细胞的吞噬和营养能力，将其转化为修复表型 修复蛋白，如血红素加氧酶1、精氨酸酶1和血管内皮生长因子α。这个 巨噬细胞和ILC2s之间的通讯已经在肾脏缺血的外周进行了研究。 再灌注损伤，但脑损伤很小，且正在老化。初步数据显示，随着年龄的增长，ILC2在 雄性和雌性小鼠的大脑与其他ILC类型的比较。此外，全球范围内男性的数量增长了好几倍。 在ILC和ILC2总数中，她们的ILC和ILC2总数多于女性。此外，男性ILC2 比女性产生更多的细胞因子，在衰老过程中具有更强的抗炎能力。最后，年轻的ILC2 可溶性因子在体外增加小胶质细胞中Nrf2和Nrf2调控的基因。这表明老化的ILC2 可能有益于炎症反应过程中的小胶质细胞极化。因此，我假设ILC2 在以下情况下，大脑的小胶质细胞极化是修复性的、M2样表型的守门人：(1)动态平衡 条件，(2)缺血性损伤，(3)衰老。在目标1中，我将研究ILC2和ILC2的转录组 可溶性因素对小胶质细胞极化的影响。在目标2中，我将确定ILC2对氧气-葡萄糖的反应 剥夺，一种缺血损伤的体外模型，以及它们的可溶性因素如何极化启动的小胶质细胞。最后， 在目标3中，我将进行概念验证实验，以探讨体外刺激的ILC2对老年人的作用 皮质内注射GFP+ILC2s过继细胞转移小胶质细胞。加在一起，我会有 确定了ILC2可溶性因子将小胶质细胞极化为修复表型的能力。这将提供 进一步探索ILC2作为中风和其他神经疾病的可能治疗平台的理论基础。