Novel ocular imaging and molecular analysis of anterior eye segment for glaucoma

青光眼眼前节的新型眼部成像和分子分析

• 批准号: 10607885
• 负责人: Tsutomu Kume
• 金额: $ 67.78万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607885
• 关键词: Adrenal Cortex Hormones; Affect; Anatomy; Angiopoietins; Animals; Anterior eyeball segment structure; Aqueous Humor; Area; Blindness; Blood Vessels; Cardiac; Cells; Characteristics; Circulation; Data; Detection; Development; Disease; Drainage procedure; Electron Microscopy; Endothelial Cells; Evaluation; Extracellular Matrix; Eye diseases; FOXC2 gene; Family; Gene Expression; Glaucoma; Goals; Height; Histologic; Human; Image; Imaging Techniques; Imaging technology; Immunohistochemistry; Impairment; Lighting; Limb structure; Lymph; Lymphatic; Lymphedema; Maintenance; Measurement; Measures; Mediating; Mesenchymal; Methodology; Methods; Mission; Molecular; Molecular Analysis; Morphology; Mus; Mutant Strains Mice; Mutation; Neural Crest; Ocular Hypertension; Optic Nerve; Optical Coherence Tomography; Pathologic; Pathway interactions; Patients; Phase; Physiologic Intraocular Pressure; Play; Public Health; Research; Resistance; Resolution; Retina; Risk Factors; Role; Scanning; Signal Pathway; Speed; Structure; Structure of sinus venosus of sclera; System; Testing; Therapeutic; Three-Dimensional Imaging; Time; Tissues; Trabecular meshwork structure; Variant; Vascular Endothelial Cell; Vascular Endothelial Growth Factor Receptor-3; Vascular System; Veins; Visible Radiation; Vision; Visual impairment; Width; Wild Type Mouse; age related; aqueous; dominant genetic mutation; early onset; experimental study; improved; in vivo; member; meter; modifiable risk; mutant; nano; nerve damage; novel; novel diagnostics; ocular imaging; older patient; optic nerve disorder; pressure; primary congenital glaucoma; single-cell RNA sequencing; therapeutically effective; transcription factor

Project Summary Glaucoma is the leading cause of irreversible blindness worldwide and is a heterogeneous group of ocular diseases characterized by optic nerve damage and impaired vision. The majority of glaucoma patients are elderly due to the age-related, optic nerve disease, whereas there are early-onset forms of glaucoma: primary congenital glaucoma and developmental glaucoma. The resistance to drainage of aqueous humor from the anterior segment of the eye into the systemic circulation leads to the elevation of intraocular pressure (IOP), and elevated IOP is the primary and only modifiable risk factor for glaucoma. Despite the current treatment options aimed at lowering IOP, there are urgent needs to develop new, more effective therapeutic approaches. Aqueous humor drainage as the conventional outflow pathway is mediated through the trabecular meshwork (TM), which is derived from the neural crest (NC) lineage, into a specialized, lymphatic-like vessel known as Schlemm's canal (SC). Recent evidence indicates that SC has specialized and unique vascular endothelial cells (ECs) and that the signaling pathways such as Angiopoietin/Tie2 are essential for the formation and maintenance of SC vasculature. The long-term goal of our group is to elucidate the fundamental mechanisms that regulate SC maintenance/function and to understand how disruption of these mechanisms leads to impaired outflow causing increased IOP in pathological settings. To image the full-circle SC and evaluate TM stiffness in mice, our group has recently developed a circular-scanning visible-light OCT (vis-OCT) system. FOXC2 is a member of the FOX (Forkhead box) transcription factor family and has critical roles in vascular development and disease. Inactivating mutations in human FOXC2 are dominantly associated with lymphedema, which is characterized by obstructed lymph drainage in the limbs, and recent evidence has shown that FOXC2 variants possess a role as putative modifiers for the development of primary congenital glaucoma. However, its role in TM and SC cells has yet to be fully investigated. The results from our preliminary experiments via vis-OCT imaging, immunohistochemistry, and single-cell RNA sequencing indicate that the NC-Foxc2-/- mutation is associated with defective TM formation, abnormal SC morphology, or the absence of the SC, a reduction in expression of PROX1, VEGFR3, and TEK in SC ECs, and elevated IOP, and that the EC-Foxc2-/- mutation results in impaired SC formation. Our central hypothesis is that novel diagnostic vis-OCT imaging techniques elucidate the significance of Foxc2 function in both NC-derived TM cells and SC ECs in SC formation/maintenance and TM stiffness. To test this hypothesis, we have two Specific Aims. First, we will develop a new balanced-detection vis-OCT to determine the anatomical influence of the Foxc2 regulatory network in SC formation/maintenance. Second, we will develop new methodologies to evaluate TM stiffness based on vis-OCT phase imaging after high-speed temporal speckle reduction to define the functional influence of the Foxc2 regulatory network in TM stiffness and ECM compositions.

项目概要 青光眼是全世界不可逆性失明的主要原因，是一种异质性眼病 以视神经损伤和视力受损为特征的疾病。大多数青光眼患者是老年人 由于与年龄相关的视神经疾病，而青光眼也有早发形式：原发性先天性 青光眼和发育性青光眼。房水从前房引流的阻力 眼睛的一部分进入体循环导致眼内压（IOP）升高，并升高 IOP 是青光眼的主要且唯一可改变的危险因素。尽管目前的治疗方案旨在 为了降低眼压，迫切需要开发新的、更有效的治疗方法。房水 引流作为传统的流出途径是通过小梁网（TM）介导的，这是 源自神经嵴 (NC) 谱系，进入称为施莱姆管的特殊淋巴管样血管 （SC）。最近的证据表明 SC 具有专门且独特的血管内皮细胞 (EC)，并且 血管生成素/Tie2等信号通路对于SC的形成和维持至关重要 脉管系统。我们小组的长期目标是阐明调节 SC 的基本机制 维护/功能并了解这些机制的破坏如何导致流出受损 病理情况下眼压升高。为了对小鼠的全圆 SC 进行成像并评估 TM 硬度，我们小组 最近开发了一种循环扫描可见光 OCT (vis-OCT) 系统。 FOXC2 是 FOX 的成员 （叉头框）转录因子家族，在血管发育和疾病中具有关键作用。失活 人类 FOXC2 突变主要与淋巴水肿相关，其特征是阻塞 四肢的淋巴引流，最近的证据表明 FOXC2 变体具有假定的作用 原发性先天性青光眼发展的调节剂。然而，其在 TM 和 SC 细胞中的作用尚未得到证实。 进行全面调查。我们通过 vis-OCT 成像、免疫组织化学进行的初步实验的结果， 单细胞 RNA 测序表明 NC-Foxc2-/- 突变与 TM 形成缺陷相关， SC 形态异常，或 SC 缺失，PROX1、VEGFR3 和 TEK 表达减少 EC-Foxc2-/- 突变导致 SC 形成受损。我们的中央 假设新的诊断 vis-OCT 成像技术阐明了 Foxc2 功能在 NC 衍生的 TM 细胞和 SC EC 在 SC 形成/维持和 TM 硬度中的作用。为了检验这个假设， 我们有两个具体目标。首先，我们将开发一种新的平衡检测 vis-OCT 来确定解剖学 Foxc2 调控网络对 SC 形成/维持的影响。第二，我们将开发新的 基于高速时间散斑后的 vis-OCT 相位成像评估 TM 刚度的方法 减少以定义 Foxc2 调节网络对 TM 硬度和 ECM 的功能影响 组成。