Immune mediated regeneration of retinal ganglion cell axons following optic nerve trauma

视神经损伤后免疫介导的视网膜神经节细胞轴突再生

基本信息

批准号：
10017241
负责人：
Benjamin M Segal
金额：
$ 35.79万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10017241
关键词：
Adoptive Transfer Apoptosis Autologous Axon Blindness Bone Marrow CD14 gene Cell Nucleus Cell Survival Cell Therapy Cell Wall Cell surface Cells Characteristics Chimera organism Clinical Colony-Stimulating Factor Receptors Crush Injury Data Development Eye Failure Generations Glaucoma Goals Granulocyte Colony-Stimulating Factor Granulocyte Colony-Stimulating Factor Receptors Growth Factor HL-60 Cells HL60 Human Immune Immunodeficient Mouse Immunomodulators In Vitro Individual Injections Injury Interleukin 4 Receptor Interleukin-4 Intervention Intravenous Kinetics Knockout Mice Lead Measures Mediating Messenger RNA Modeling Morphology Mus Myelogenous Natural regeneration Nerve Crush Nerve Fibers Nerve Regeneration Neurons Neutrophilic Infiltrate Nuclear Optic Nerve Optic Nerve Injuries Optic Neuritis Output Pathologic Patients Phenotype Property Proteins Protocols documentation Receptor Signaling Recombinants Research Retinal Ganglion Cells Role Secondary to Signal Transduction Signaling Molecule Source Testing Therapeutic Therapeutic Uses Transcript Trauma Tretinoin Up-Regulation Visual Vitreous body structure Zymosan arginase axon regeneration axonopathy conditional knockout dectin 1 experimental study gain of function immunoregulation in vivo innovation neutrophil novel novel therapeutic intervention novel therapeutics optic nerve disorder prevent receptor receptor expression regenerative repaired response retinal ganglion cell regeneration

项目摘要

Axonopathy is an early and prominent pathological feature of glaucoma, optic neuritis and traumatic optic nerve injury. Permanent loss of vision in all of these conditions is secondary, in large part, to a failure of retinal ganglion cells (RGC), the output neurons of the optic nerve, to survive and regenerate their axons. There is a dire need to develop novel therapeutic interventions that overcome barriers to repair in the eye, promote RGC survival and RGC axonal regrowth, thereby mitigating, or even reversing, visual loss. In this proposal we investigate a novel subset of neutrophils that accumulate in the vitreous body following intraocular (i.o.) injection of mice with the fungal cell wall extract, zymosan, and are associated with the regrowth of severed RGC axons. In preliminary studies we have demonstrated that adoptive transfer of zymosan-elicited neutrophils directly into the vitreous of mice with optic nerve crush (ONC) injury is sufficient to rescue RGC and stimulate RGC axon regrowth. These neutrophils are characterized by ring-form nuclei and the cell surface phenotype CD14+Ly6Glow. They express high levels of transcripts for arginase-1 and CD206. Our major goals are to elucidate the factors that drive the differentiation of reparative neutrophils and develop protocols to generate them in vitro for therapeutic application in individuals with optic neuropathy. In Aim 1 we will test our hypothesis that IL-4 and granulocyte-colony stimulating factor (G-CSF) act synergistically to drive the differentiation of pro-regenerative neutrophils in vivo in mice subjected to i.o. zymosan injection and ONC injury. A role of IL-4 was suggested by our finding that CD14+Ly6Glow neutrophils express high levels of IL-4 signaling molecules, and IL-4 protein is produced in the vitreous following i.o. administration of zymosan. G- CSF is also upregulated in the vitreous and it was recently shown to induce IL-4 receptor expression on neutrophils. We will determine the kinetics and cellular source of IL-4, IL-4 receptor chains, G-CSF and G-CSF receptor in zymosan injected eyes. Loss and gain of function experiments, using a panel of conditional knock- out mice and bone marrow chimeras, will be performed to assess the roles of IL-4 and G-CSF signaling in the development of CD14+Ly6Glow neutrophils, RGC survival and axonal regeneration following i.o. zymosan injection and ONC. This research could ultimately lead to the development of novel, or the repurposing of established, immunomodulators to promote the differentiation and expansion of neuroregenerative neutrophils in patients with optic neuropathy secondary to glaucoma, optic neuritis or trauma. Our exploratory experiments have shown that murine bone marrow neutrophils acquire characteristics of zymosan-elicited CD14+Ly6Glow neutrophils, including the ability to drive axonal regeneration, following in vitro polarization with IL-4 and G- CSF. The overall goal of Aim 2 is to optimize protocols for the generation of pro-regenerative neutrophils from murine bone marrow precursors ex vivo. The goal of Aim 3 is to assess the neuroregenerative potential of IL-4 modulated human neutrophils that could, potentially, be re-infused into patients with optic neuropathy, as an autologous cell therapy.

轴突病是青光眼、视神经炎和外伤性视神经的早期、突出的病理特征。神经损伤。所有这些情况下的永久性视力丧失在很大程度上都是继发于视网膜功能衰竭的神经节细胞（RGC）是视神经的输出神经元，能够生存并再生轴突。有一个迫切需要开发新的治疗干预措施来克服眼睛修复障碍，促进 RGC 存活和 RGC 轴突再生，从而减轻甚至逆转视力丧失。在这个提案中我们研究眼内（i.o.）后在玻璃体内积聚的中性粒细胞的新亚群给小鼠注射真菌细胞壁提取物酵母聚糖，并与被切断的细胞壁的再生有关 RGC 轴突。在初步研究中，我们已经证明酵母聚糖的过继转移引起将中性粒细胞直接注入视神经挤压 (ONC) 损伤小鼠的玻璃体足以挽救 RGC 刺激 RGC 轴突再生。这些中性粒细胞的特征是环形核和细胞表面表型 CD14+Ly6Glow。它们表达高水平的精氨酸酶 1 和 CD206 转录本。我们的主要目标旨在阐明驱动修复性中性粒细胞分化的因素并制定方案在体外生成它们用于视神经病患者的治疗应用。在目标 1 中，我们将测试我们的假设 IL-4 和粒细胞集落刺激因子 (G-CSF) 协同作用以驱动小鼠体内促再生中性粒细胞的分化酵母聚糖注射液和ONC 受伤。我们发现 CD14+Ly6Glow 中性粒细胞表达高水平的 IL-4，表明了 IL-4 的作用信号分子，IL-4 蛋白在注射后在玻璃体中产生。酵母聚糖的施用。 G- CSF 在玻璃体中的表达也上调，最近显示它可以诱导玻璃体中 IL-4 受体的表达。中性粒细胞。我们将确定 IL-4、IL-4 受体链、G-CSF 和 G-CSF 的动力学和细胞来源注射酵母聚糖的眼睛中的受体。功能实验的损失和增益，使用一组条件敲击小鼠和骨髓嵌合体，将被用来评估 IL-4 和 G-CSF 信号在注射后 CD14+Ly6Glow 中性粒细胞的发育、RGC 存活和轴突再生酵母聚糖注射和ONC。这项研究最终可能会导致小说的发展，或者重新利用已建立免疫调节剂促进神经再生中性粒细胞的分化和扩张继发于青光眼、视神经炎或外伤的视神经病变患者。我们的探索性实验已表明小鼠骨髓中性粒细胞获得酵母聚糖引发的 CD14+Ly6Glow 的特征中性粒细胞，包括在体外用 IL-4 和 G- 极化后驱动轴突再生的能力脑脊液。目标 2 的总体目标是优化从细胞中生成促再生中性粒细胞的方案小鼠骨髓前体离体。目标 3 的目标是评估 IL-4 的神经再生潜力调节的人类中性粒细胞有可能被重新输注到视神经病变患者体内，作为一种治疗方法自体细胞疗法。