Innate Immunomodulation of Retinal Vascular Development

视网膜血管发育的先天免疫调节

• 批准号: 10924832
• 负责人: Gopalan Gnanaguru
• 金额: $ 44.5万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10924832
• 关键词: Ablation; Address; Adult; Affect; Animals; Astrocytes; Biological Assay; Birth; Blindness; Blood Vessels; C3AR1 gene; CCL2 gene; CD11c Antigens; Cell Separation; Cells; Cellular Morphology; Complement; Complement 3a; Complement Activation; Complement Receptor; Complex; Custom; Data; Development; Developmental Process; Distal; Embryo; Endothelial Cells; Endothelium; Exhibits; Future; Gene Expression; Goals; Growth; Immune; Immunoassay; In Situ Hybridization; In Vitro; Innate Immune System; Knockout Mice; Knowledge; Ligands; Macrophage-1 Antigen; Microglia; Migration Assay; Molecular; Mus; Natural Immunity; Neonatal; Neuroglia; Optic Disk; Optic Nerve; Pathology; Pattern; Phagocytosis; Phase; Physiological; Population; Process; Proliferating; Proteins; RNA; Regulation; Reporter; Reporting; Retina; Retinopathy of Prematurity; Reverse Transcriptase Polymerase Chain Reaction; Role; Shapes; Signal Transduction; Testing; Therapeutic; Transcript; Vascular Diseases; Vision; blood vessel development; cell motility; cell type; chemokine; chemokine receptor; design; gene complementation; immune activation; immunoregulation; innovation; migration; mutant; neovasculature; new therapeutic target; novel; novel therapeutic intervention; postnatal; receptor; recruit; retina blood vessel structure; targeted treatment; transcriptome sequencing

Project Summary Retinal blood vessel formation is a highly regulated process that requires multi-cellular crosstalk and interactions. Due to the complexity of retinal vascular development, there are several critical knowledge gaps that need to be addressed. For example, the key glial cell type that regulate retinal vascular development are astrocytes, which lay a template for blood vessel formation. Endothelial cells migrate over the spatially organized astrocytic template to form superficial blood vessels that give rise to three interconnected vascular layers in the mature retina. Disruption of the astrocytic template during development or loss of astrocyte association with blood vessels in adulthood can be detrimental to vascular growth, integrity, and function. The underlying cellular and molecular signaling mechanisms that regulate astrocyte spatial patterning and subsequent organized blood vessel formation during development remain incompletely understood. Elucidating the molecular mechanisms that govern blood vessel development and function is necessary to identify more targeted therapeutic strategies for blinding retinal vascular pathologies, and to identify which critical developmental processes should not be targeted in some contexts, for example retinopathy of prematurity. Our major goal is to delineate the signaling mechanisms that regulate astrocyte template spatial arrangement and vascular network formation. Our preliminary findings strongly support the rationale for the present study. Our RNA-seq data reveal that specific chemokine and complement gene expression levels are elevated during retinal vascular development. Intriguingly, deletion of one of the affected chemokine receptors disrupts astrocyte template formation and microglial recruitment and distribution. On the other hand, deletion of complement components results in an aberrantly dense astrocytic template and dysmorphic excessive tip cell formation. Based on these novel findings, we hypothesize that microglial chemokine signaling and complement activation are critical for normal retinal vascular development. We will refute or validate our hypothesis in the following two Specific Aims: Aim 1: To determine if chemokine signaling recruits microglia to modulate astrocyte template assembly. Aim 2: To define the role of complement receptors in astrocyte template and vascular network formation. We will utilize innovative multiplex RNA/protein based assays, novel ex vivo migration assays, unique reporter mice, and cre/lox animals for cell-specific deletion. We expect that successful completion of the proposed studies will identify novel roles for the innate immune system in regulating highly complex retinal vascular developmental processes. Moreover, further elucidating physiological regulatory mechanisms of vascular development, will also identify future putative therapeutic strategies for retinal vascular pathologies.

项目摘要 视网膜血管形成是一个高度调节的过程，需要多细胞串扰和 互动。由于视网膜血管发育的复杂性，存在几个关键的知识差距 需要解决。例如，调节视网膜血管发育的关键神经胶质细胞类型是 星形胶质细胞为血管形成铺设模板。内皮细胞在空间上迁移 有组织的星形细胞模板形成浅表血管，产生三个相互连接的血管 成熟视网膜中的层。星形胶质细胞开发或丢失的星形细胞模板的破坏 在成年期与血管的关联可能不利于血管生长，完整性和功能。这 基础的细胞和分子信号传导机制，调节星形胶质细胞空间模式和 随后在发育过程中的有组织的血管形成尚未完全理解。阐明 为了识别更多的血管发育和功能的分子机制对于识别更多 针对视网膜血管病理盲目的有针对性的治疗策略，并确定哪种关键 发展过程不应在某些情况下针对性，例如早产性视网膜病变。我们的 主要目标是描述调节星形胶质细胞模板空间布置和的信号传导机制 血管网络形成。 我们的初步发现强烈支持本研究的基本原理。我们的RNA-seq数据表明 在视网膜血管发育过程中，特定的趋化因子和补体基因表达水平升高。 有趣的是，其中一种受影响的趋化因子受体的删除破坏了星形胶质细胞模板的形成和 小胶质招募和分布。另一方面，补体组件的删除导致 异常致密的星形胶质细胞模板和畸形过多的尖端细胞形成。基于这些小说 调查结果，我们假设小胶质细胞趋化因子信号传导和补体激活对于正常 视网膜血管发育。我们将在以下两个具体目标中驳斥或验证我们的假设：目标 1：确定趋化因子信号是否募集小胶质细胞调节星形胶质细胞模板组件。目标2：到 定义补体受体在星形胶质细胞模板和血管网络形成中的作用。我们将利用 创新的多重RNA/基于蛋白质的测定，新型的离体迁移测定，独特的记者小鼠和 Cre/Lox动物用于细胞特异性缺失。我们希望成功完成拟议的研究将 确定先天免疫系统在调节高度复杂的视网膜血管发育中的新作用 过程。此外，进一步阐明了血管发育的生理调节机制，将 还可以确定视网膜病理病理的未来假定治疗策略。

项目成果

Retinal Injury Activates Complement Expression in Müller Cells Leading to Neuroinflammation and Photoreceptor Cell Death.

• DOI: 10.3390/cells12131754
• 发表时间: 2023-06-30
• 期刊: CELLS
• 影响因子: 6
• 作者: Tabor, Steven J. J.;Yuda, Kentaro;Deck, Jonathan;Gnanaguru, Gopalan;Connor, Kip M. M.
• 通讯作者: Connor, Kip M. M.