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，视神经炎和创伤性视神经的早期且突出的病理特征神经损伤。在所有这些条件下，永久视力丧失在很大程度上都是视网膜失败的神经节细胞（RGC）是视神经的输出神经元，以存活并再生其轴突。有一个迫切需要开发新的治疗干预措施，以克服眼睛修复的障碍，促进RGC 生存和RGC轴突再生，从而减轻甚至逆转视觉丧失。在这个建议中，我们研究一个新的嗜中性粒细胞的子集，该子集在眼内（I.O.）后积聚在玻璃体体内用真菌细胞壁提取物，Zymosan注射小鼠，与切断的再生有关 RGC轴突。在初步研究中，我们已经证明了Zymosan-PELITISET的收养转移嗜中性粒细胞直接进入视神经压伤（ONC）损伤的小鼠玻璃体足以挽救RGC，并且刺激RGC轴突再生。这些中性粒细胞以环形核和细胞表面为特征表型CD14+Ly6glow。它们表达精氨酸酶-1和CD206的高水平转录本。我们的主要目标阐明驱动差异性中性粒细胞区分的因素，并为在患有视神经病理的个体中进行治疗应用，在体外生成它们。在AIM 1中，我们将测试我们的假设IL-4和粒细胞 - 粘合剂刺激因子（G-CSF）协同行动以驱动在i.o的小鼠中体内促嗜中性粒细胞的分化Zymosan注入和ONC 受伤。我们发现CD14+Ly6glow中性粒细胞表达高水平的IL-4，提出了IL-4的作用信号分子和IL-4蛋白在I.O.后面的玻璃体中产生。 Zymosan的给药。 g- CSF在玻璃体中也被上调，最近证明它会诱导IL-4受体表达中性粒细胞。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 Zymosan注射眼睛的受体。功能实验的丢失和增益，使用一组条件性敲门板将进行鼠标和骨髓嵌合体，以评估IL-4和G-CSF信号的作用 i.o. Zymosan 注射和ONC。这项研究最终可能导致小说的发展，或者重新利用已建立的免疫调节剂，以促进神经增生中性粒细胞的分化和扩展继发于青光眼，视神经炎或创伤的患有神经病的患者中。我们的探索实验已经表明，鼠骨髓嗜中性粒细胞获得了Zymosan引诱的CD14+ly6glow的特征嗜中性粒细胞，包括驱动轴突再生的能力。 CSF。 AIM 2的总体目标是优化为从鼠骨髓前体离体。 AIM 3的目的是评估IL-4的神经加快潜力调节的人类嗜中性粒细胞可能会被重新注入视神经病患者，作为一种自体细胞疗法。