Mechanisms of microglial neuroinflammatory response in glaucoma

青光眼小胶质细胞神经炎症反应机制

基本信息

批准号：
10717247
负责人：
Milica Margeta
金额：
$ 49.25万
依托单位：
SCHEPENS EYE RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10717247
关键词：
Animal Model Animals Apolipoprotein E Apoptotic Aqueous Humor Autoimmune Autopsy Binding Blindness Brain Cause of Death Cell Culture Techniques Cell Death Cell Survival Cells Characteristics Complement Data Data Set Demyelinations Development Disease Eye Galectin 3 Genes Genetic Glaucoma Goals Human Immune Immunoglobulin Fragments In Vitro Inflammasome Inflammation Inflammatory Inflammatory Response Injections Interleukins Macrophage Mediating Mediator Membrane Mental Depression Microglia Microspheres Modeling Molecular Molecular Profiling Morbidity - disease rate Mus Myeloid Cells NR0B2 gene Nerve Degeneration Nervous System Neurodegenerative Disorders Neurons Optic Nerve PIK3CG gene PTPN11 gene Pathogenesis Patients Phenotype Phosphoric Monoester Hydrolases Physiologic Intraocular Pressure Play Prevalence Regulation Reporter Research Retina Retinal Ganglion Cells Risk Factors Role Signal Pathway Signal Transduction Therapeutic Up-Regulation Visual impairment Work apolipoprotein E-3 blind cell injury common treatment conditional knockout cytokine efficacy evaluation glial activation high intraocular pressure in vivo modifiable risk monocyte mouse model neuroinflammation neuron loss neuroprotection neutralizing antibody novel peripheral nerve regeneration pharmacologic phosphatidylserine receptor prevent receptor recruit response retinal ganglion cell degeneration sensor transcriptome sequencing translational approach

项目摘要

The goal of this proposal is to elucidate the role of microglia, the resident immune cells of the retina and the brain, in the pathogenesis of glaucoma. Glaucoma is the leading cause of irreversible blindness worldwide, whose hallmark is progressive loss of retinal ganglion cells (RGCs). In our prior work we have found that microglia in glaucoma upregulate a disease-associated molecular signature that is shared with brain neurodegenerations, and that genetic or pharmacologic targeting of molecules in this signaling pathway can prevent RGC loss in a mouse model of glaucoma. Furthermore, we have found that microglia activated by engulfment of apoptotic neurons can subsequently cause more RGC loss in the absence of high intraocular pressure (IOP), demonstrating that microglia can propagate neuronal damage in glaucoma. Using the RNAseq datasets from these studies, we have now identified the transmembrane receptor CD300lf as a key mediator of microglial inflammatory response in glaucoma. CD300lf is a phosphatidylserine receptor with dual activating and inhibitory capacity that can bind to damaged and apoptotic cells and, in the brain, is expressed only by myeloid cells. Our preliminary data demonstrate that 1) CD300lf is upregulated in retinal microglia from two mouse glaucoma models; 2) CD300lf–/– animals are protected from RGC loss in the microbead glaucoma model despite elevated IOP; and 3) unlike wildtype microglia, CD300lf–/– microglia do not upregulate inflammatory cytokines, including Il-1, in response to damaged/apoptotic neurons in vivo. Based on these data, we hypothesize that CD300lf is a key regulator of the microglial inflammatory response that exacerbates RGC loss in glaucoma. We will address this hypothesis with the following Specific Aims: 1. Determine if microglia are the key myeloid cell subpopulation that mediates CD300lf effect in glaucoma. In this aim, we will use CD300lffl/fl conditional knockouts to identify whether CD300lf harms RGCs by acting in microglia or other myeloid cell subpopulations (monocytes, border-associated macrophages). 2: Investigate the effect of targeting CD300lf and its binding partners on inflammasome signaling. We will evaluate the inflammatory response and the inflammasome function in myeloid cells deficient in CD300lf or its binding partners in cell culture and in vivo. 3: Investigate the therapeutic potential of targeting CD300lf in glaucoma. We will assess the efficacy of CD300lf neutralizing antibody fragment in protecting RGCs in a mouse model of glaucoma and evaluate if CD300LF is upregulated in human postmortem eyes with glaucoma. The research outlined in this proposal will define the role of CD300lf signaling in microglia in glaucoma, with the ultimate goal of developing novel neuroprotective treatments for this common blinding disease.

该提案的目标是阐明小胶质细胞、视网膜常驻免疫细胞和视网膜免疫细胞的作用。青光眼的发病机制中，大脑是全球不可逆性失明的主要原因。其标志是视网膜神经节细胞（RGC）的逐渐丧失。在我们之前的工作中，我们发现了这一点。青光眼中的小胶质细胞上调与大脑共享的与疾病相关的分子特征神经退行性疾病，并且该信号通路中分子的遗传或药理学靶向可以防止青光眼小鼠模型中的 RGC 损失此外，我们发现小胶质细胞被激活。在没有高眼压的情况下，凋亡神经元的吞噬随后会导致更多的 RGC 损失。使用 RNAseq 证明小胶质细胞可以传播青光眼中的神经元损伤。根据这些研究的数据集，我们现在已经确定跨膜受体 CD300lf 是 CD300lf 是一种具有双重激活作用的磷脂酰丝氨酸受体。能够与受损细胞和凋亡细胞结合的抑制能力，在大脑中仅由髓系细胞表达我们的初步数据表明 1) CD300lf 在两只小鼠的视网膜小胶质细胞中上调。青光眼模型；2) CD300lf–/– 动物在微珠青光眼模型中免受 RGC 损失，尽管眼压升高；3) 与野生型小胶质细胞不同，CD300lf–/– 小胶质细胞不会上调炎症细胞因子，包括 Il-1，针对体内受损/凋亡神经元的反应，基于这些数据，我们勇敢地面对这一点。 CD300lf 是小胶质细胞炎症反应的关键调节因子，可加剧青光眼中 RGC 的损失。将通过以下具体目标来解决这一假设： 1. 确定小胶质细胞是否是介导 CD300lf 效应的关键骨髓细胞亚群在此目标中，我们将使用 CD300lfl/fl 条件敲除来确定 CD300lf 是否通过以下方式损害 RGC。作用于小胶质细胞或其他骨髓细胞亚群（单核细胞、边界相关巨噬细胞）。图 2：研究靶向 CD300lf 及其结合伴侣对炎性体信号转导的影响。将评估缺乏 CD300lf 或的骨髓细胞的炎症反应和炎症小体功能它在细胞培养和体内的结合伙伴。 3：研究靶向 CD300lf 在青光眼中的治疗潜力我们将评估其疗效。 CD300lf 中和抗体片段在青光眼小鼠模型中保护 RGC 的作用并评估是否 CD300LF 在人类死后患有青光眼的眼睛中表达上调。本提案中概述的研究将定义 CD300lf 信号在青光眼小胶质细胞中的作用，最终目标是开发针对这种常见致盲疾病的新型神经保护疗法。