The role of optic nerve lamina region stem cells in age-related optic nerve disease

视神经板区域干细胞在年龄相关性视神经疾病中的作用

基本信息

批准号：
10443202
负责人：
STEVEN L BERNSTEIN
金额：
$ 42.73万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10443202
关键词：
Acute Age Aging Anterior Anterior Ischemic Optic Neuropathy Apoptosis Astrocytes Axon Biological Blindness Blood Vessels Cell Culture Techniques Cell Death Cell Survival Cells Cellular Stress Cessation of life Coupling Defect Disease Disease Resistance Dissection Distal Eye Ganciclovir Glaucoma Growth Growth Factor Health Human Individual Mediating Membrane MicroRNAs Modeling Molecular Mus Natural regeneration Neurosphere Non-Rodent Model Nuclear Pore Complex Optic Disk Optic Nerve Patients Phagocytes Physiologic Intraocular Pressure Predisposition Prevalence Primary Open Angle Glaucoma Process Property Proteins Rattus Resistance Retina Retinal Ganglion Cells Rodent Rodent Model Role Stress Supporting Cell Techniques Thinking Thymidine Kinase Transgenic Mice Transgenic Model Vesicle Vision age related aged axonopathy biomarker panel deep sequencing effective therapy enhancing factor exosome extracellular vesicles improved improved outcome in vivo injury recovery innovation myelination nanovesicle nerve stem cell nestin protein neuroprotection nonhuman primate optic nerve disorder postnatal stem cell biomarkers stem cells

项目摘要

The most common age-associated optic nerve (ON)-related causes of vision loss are non-arteritic anterior ischemic optic neuropathy (NAION) and primary open-angle glaucoma (POAG). Optic nerve head (ONH) defects contribute to NAION and POAG susceptibility, but the mechanisms responsible for this are incompletely understood, contributing to a lack of effective treatments. My lab recently discovered that the optic nerve-laminar region (ONLR) within the ONH, contains a CNS neural stem cell/neural progenitor cell (NSC/NPC) niche which is depleted during aging. Increased CNS-NPC activity has been shown to improve baseline CNS activity in aged mice, and NPC depletion impedes CNS recovery following injury. NPCs secrete extracellular vesicles (‘exosomes’) that mediate many of the positive effects of CNS- NPCs. We find that administering human ONLR-NPC-secreted exosomes enhance RGC survival ex vivo and stimulates RGC-neuritigenesis. Depleting ONLR-NPCs in a mouse transgenic model increases markers of RGC stress, using an RGC stress-marker panel. I hypothesize that ONLR-NPCs support RGC survival, and they do this in part by vesicle secretion. I predict their loss increases RGC stress and susceptibility to death after axonal ischemic stress, and supplementing RGCs with ONLR-NPC-extracellular vesicles will enhance RGC survival after axonal stress. We will prove this with two rodent species and two specific aims. Specific aim 1: Demonstrate that mouse ONLR-NPC loss results in RGC stress and increases RGC death in ON disease. We will couple a mouse transgenic model enabling selective ONLR-NPC depletion, the rodent model of NAION rNAION model, and stereology (statistically robust cell quantification). We will utilize molecular and cell biological techniques for identifying RGC cell stress and apoptosis. We will then: A) Determine whether acute ONLR-NPC depletion results in RGC stress, via stress marker analysis and B) Whether this depletion enhances RGC death after rNAION-induced RGC ischemic axonal stress. I predict increased RGC stress, demonstrable by increased RGC-pJun expression and increased RGC loss. Specific aim 2. Confirm that rat ONLR-NPC ‘exosomes’ protect RGCs in culture and in vivo during ischemic ON stress. We will isolate rat ONLR-NPC secreted vesicles and confirm their ability to enhance RGC survival, by: A) Administering rat ONLR-NPC vesicles and their dissociated components to cultured rat RGCs, we will quantify their ability to enhance RGC survival and neuritigenesis. I predict improved RGC survival and increased neuritigenesis. B) Using the rNAION model, we will intravitreally inject rat ONLR-NPC derived exosomes and their components and determine whether these vesicles improve RGC survival and reduce stress-related markers such as CRF and pJun. I predict reduced stress levels and improved long-term RGC survival. Our innovative approach will identify the factors that contribute to RGC stress resistance and generate new, improved approaches to treatment of optic nerve diseases.

与年龄相关的视神经 (ON) 相关的视力丧失最常见原因是非动脉炎性的前部缺血性视神经病变（NAION）和原发性开角型青光眼（POAG）。 (ONH) 缺陷会导致 NAION 和 POAG 易感性，但造成这种情况的机制是不完全了解，导致缺乏有效的治疗方法，我的实验室最近发现光学器件。 ONH 内的神经层区域 (ONLR)，包含 CNS 神经干细胞/神经祖细胞 (NSC/NPC) 生态位在衰老过程中被耗尽，中枢神经系统-NPC 活性的增加已被证明可以改善。老年小鼠的基线 CNS 活动和 NPC 耗竭会阻碍损伤后 CNS 的恢复。 NPC 分泌细胞外囊泡（“外泌体”），介导中枢神经系统的许多积极作用我们发现，施用人类 ONLR-NPC 分泌的外泌体可增强 RGC 的离体存活率，并且刺激 RGC 神经炎发生。在小鼠转基因模型中消耗 ONLR-NPC 会增加标记物。 RGC 压力，使用 RGC 压力标记组，我发现 ONLR-NPC 支持 RGC 存活，并且他们部分是通过囊泡分泌来做到这一点的，我预测他们会增加 RGC 压力的损失和死亡的可能性。轴突缺血应激后，用 ONLR-NPC-细胞外囊泡补充 RGC 将增强轴突应激后 RGC 的存活率我们将用两种啮齿类动物和两个特定目标来证明这一点。具体目标 1：证明小鼠 ONLR-NPC 缺失会导致 RGC 应激并增加 ON 疾病中的 RGC 死亡我们将结合小鼠转基因模型，实现选择性 ONLR-NPC。耗竭、NAION rNAION 模型的啮齿动物模型和体视学（统计稳健的细胞定量）。我们将利用分子和细胞生物学技术来识别 RGC 细胞应激和细胞凋亡。然后：A) 通过应激标记分析和确定急性 ONLR-NPC 耗竭是否会导致 RGC 应激 B) 我预测 rNAION 诱导的 RGC 缺血性轴突应激后这种消耗是否会增强 RGC 死亡。 RGC 应激增加，可通过 RGC-pJun 表达增加和 RGC 损失增加来证明。具体目标 2. 确认大鼠 ONLR-NPC“外泌体”在培养和体内保护 RGC 我们将分离大鼠ONLR-NPC分泌囊泡并确认其增强能力。 RGC 存活，通过：A) 将大鼠 ONLR-NPC 囊泡及其解离成分施用于培养的大鼠 RGC，我们将量化它们增强 RGC 存活和神经炎发生的能力。我预测 RGC 会得到改善。 B) 使用 rNAION 模型，我们将玻璃体内注射大鼠 ONLR-NPC。衍生的外泌体及其成分，并确定这些囊泡是否改善 RGC 存活率和减少与压力相关的标记物，例如 CRF 和 pJun，我预测压力水平会降低，长期会有所改善。我们的创新方法将确定导致 RGC 抵抗和抵抗的因素。产生新的、改进的视神经疾病治疗方法。