Innate immune system regulation of retinal regeneration

先天免疫系统对视网膜再生的调节

基本信息

批准号：
10707048
负责人：
JEFFREY MUMM
金额：
$ 44.1万
依托单位：
JOHNS HOPKINS UNIVERSITY
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 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707048
关键词：
Ablation Acute Address Adult Bioinformatics CRISPR/Cas technology Cell Death Cells Cone Data Development Disease Fatty acid glycerol esters Feedback Fishes Genes Genetic Genetic screening method Goals Histology Human Image Immune Immune system Immunosuppression In Vitro Injury Innate Immune System Kinetics Label Larva Link Mammals Methodology Methods Microglia Modeling Molecular Natural regeneration Nature Outcome Patients Photoreceptors Play Process Publishing Regenerative capacity Regulation Resources Retina Retinal Cone Retinal Ganglion Cells Role Shapes Specificity System Testing Therapeutic Time Titrations Transgenic Organisms Vision Vision Disorders Zebrafish cell type experimental study gene network glial activation immunoregulation improved in vivo in vivo imaging insight mathematical model novel progenitor programs regeneration potential regenerative regenerative therapy repaired retinal neuron retinal progenitor cell retinal regeneration sight restoration stem cell niche stem cell proliferation stem cells transcriptomics

项目摘要

PROJECT SUMMARY Degenerative vision disorders are caused by the loss of retinal cells. In zebrafish, retinal Müller glial (MG) act as stem cells, generating MG-derived progenitor cells (MGPCs) which replace lost retinal cells. Mammalian MG/MGPCs can also produce new retinal neurons, however, proliferative capacity is limited and disease- relevant cell types (e.g., photoreceptors) are rarely generated. Our goal is to understand why the regenerative potential of MG/MGPCs are limited in mammals in comparison to a regenerative species (i.e. zebrafish). In zebrafish, widespread retinal cell loss triggers “developmental” regeneration, producing all major retinal cell classes. Conversely, selective retinal cell loss results in a “fate-biased” process where MGPCs give rise to the lost cell type. How MG sense the extent of loss, however, is unknown. As the immune system plays critical roles during retinal regeneration, we examined the impact of microglia reactivity on MG activation in zebrafish. We observed that microglia are required for MG stem cell activation and that immunosuppression can inhibit or enhance regeneration kinetics depending on the treatment timing. We hypothesize that microglia reactivity levels scale to the extent, duration, and/or specificity of retinal cell loss in order to coordinate MG activation, MGPC proliferation rates and fat decisions to the type of injury incurred. To test our hypothesis, here we propose to combine an improved targeted cell ablation system enabling titratable, sustainable, and selective retinal cell loss with resources/methods for labeling of immune cell types, intravital timelapse imaging, lineage tracing, and single cell transcriptomics. Specifically we will define how the extent (Aim 1), the duration (Aim 2) and specificity (Aim 3) of cone photoreceptor or retinal ganglion cell loss impact immune cell reactivity and retinal regeneration. These experiments will serve to define roles specific immune cell subtypes play in shaping MGPC proliferation and cell fates during regeneration in zebrafish.

项目概要退行性视力障碍是由斑马鱼视网膜细胞损失引起的，视网膜穆勒胶质细胞 (MG) 起作用。作为干细胞，产生 MG 衍生的祖细胞 (MGPC)，替代丢失的哺乳动物视网膜细胞。 MG/MGPCs也可以产生新的视网膜神经元，然而，增殖能力有限且疾病- 相关细胞类型（例如光感受器）很少产生，我们的目标是了解为什么可以再生。与再生物种（即斑马鱼）相比，哺乳动物中 MG/MGPC 的潜力有限。在斑马鱼中，广泛的视网膜细胞丢失触发“发育”再生，产生所有主要视网膜细胞线下、选择性视网膜细胞丢失会导致“命运偏向”的过程，其中 MGPC 会产生“命运偏向”的过程。然而，MG 如何感知丢失的程度尚不清楚，因为免疫系统起着至关重要的作用。为了研究斑马鱼视网膜再生过程中的作用，我们研究了小胶质细胞反应性对 MG 激活的影响。我们观察到小胶质细胞是 MG 干细胞激活所必需的，并且免疫抑制可以抑制或抑制我们捕捉到了小胶质细胞的反应性，从而增强了再生动力学。水平根据视网膜细胞损失的程度、持续时间和/或特异性进行缩放，以协调 MG 激活、MGPC 增殖率和脂肪决定发生损伤的类型。假设，在这里我们建议结合改进的靶向细胞消融系统，实现可滴定，通过标记免疫细胞类型、活体细胞的资源/方法，实现可持续的、选择性的视网膜细胞损失具体来说，我们将定义延时成像、谱系追踪和单细胞转录组学的程度。（目标 1）、持续时间（目标 2）和特异性（目标 3）对视锥细胞或视网膜神经节细胞损失的影响免疫细胞反应性和视网膜再生将有助于确定特异性免疫的作用。细胞亚型在斑马鱼再生过程中影响 MGPC 增殖和细胞命运。