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

• 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
• 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

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.

轴索病是青光眼、视神经炎和外伤性视神经病变的早期和突出的病理特征 神经损伤在所有这些情况下，永久性视力丧失在很大程度上是继发于视网膜功能障碍。 神经节细胞（RGC），视神经的输出神经元，存活并再生其轴突。有一个 迫切需要开发新的治疗干预措施，克服眼睛修复障碍，促进RGC 存活和RGC轴突再生，从而减轻或甚至逆转视力丧失。在本提案中，我们 研究眼内（i.o.） 注射小鼠的真菌细胞壁提取物，酵母聚糖，并与再生的切断 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了 中性粒细胞直接进入具有视神经挤压（ONC）损伤的小鼠的玻璃体中足以拯救RGC， 刺激RGC轴突再生。这些中性粒细胞的特点是环形核和细胞表面 表型CD 14 + Ly 6 Glow。它们表达高水平的CD 206和CD 206的转录物。我们的主要目标 是为了阐明驱动修复性中性粒细胞分化的因素，并制定方案， 在体外产生它们用于视神经病变个体的治疗应用。在目标1中，我们将测试 假设IL-4和粒细胞集落刺激因子（G-CSF）协同作用以驱动 在i.o.酵母多糖注射液与ONC 损伤我们发现CD 14 + Ly 6 Glow中性粒细胞表达高水平的IL-4，这表明IL-4的作用 i.o.后，玻璃体中产生IL-4蛋白。酵母聚糖的施用。G-的 CSF在玻璃体中也上调，最近显示其诱导IL-4受体在玻璃体上的表达。 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 酵母多糖注射的眼睛中的受体。功能实验的损失和增益，使用一组有条件的敲击- 小鼠和骨髓嵌合体，将进行评估IL-4和G-CSF信号转导中的作用， i.o.后CD 14 + Ly 6 Glow中性粒细胞的发育、RGC存活和轴突再生。酵母聚糖 注射和ONC。这项研究可能最终导致小说的发展，或重新利用 已建立的免疫调节剂，以促进神经再生中性粒细胞的分化和扩增 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 已经表明小鼠骨髓嗜中性粒细胞获得酵母多糖诱导的CD 14 + Ly 6 Glow的特征 中性粒细胞，包括驱动轴突再生的能力，在体外极化与IL-4和G- 脑脊液。目标2的总体目标是优化从骨髓中产生促再生中性粒细胞的方案。 离体鼠骨髓前体。目的3的目标是评估IL-4的神经再生潜力 调节的人中性粒细胞，可以潜在地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