The role of LTBP2 in glaucoma

LTBP2在青光眼中的作用

基本信息

批准号：
10608873
负责人：
Gillian Jane McLellan
金额：
$ 23.33万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
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：
2023-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10608873
关键词：
Actins Address Adult Affect Age Angiography Animal Model Animals Anterior Aqueous Humor Binding Proteins Biological Biological Assay Biology Birth Blindness Catalogs Cell Culture Techniques Cell Proliferation Cell-Matrix Junction Cells Cellular biology Childhood Ciliary epithelium Clinical Complex Cytoskeleton Data Deposition Development Distal Ensure Equilibrium Extracellular Matrix Eye Family Felidae Felis catus Fibrosis Fluorescence Microscopy Focal Adhesions Foundations Functional disorder Future Gene Expression Genes Genetic Models Glaucoma Goals Heterozygote Homeostasis Homologous Gene Human Hydrophthalmos Image In Situ In Vitro Incubated Inherited Knock-out Knowledge Label Lasers Life Light MFAP1 gene Mediating Microfibrils Microfilaments Microscopy Modeling Molecular Morphology Mutation Open-Angle Glaucoma Pathogenesis Pathologic Pathology Pathway interactions Patients Persons Phenotype Physiologic Intraocular Pressure Physiological Physiology Play Process Production Proliferating Protein Isoforms Proteins Publishing Recombinants Regulation Reporting Resistance Role Signal Transduction Small Interfering RNA Stains Structure Testing Therapeutic Therapeutic Intervention Thick Filament Third Pregnancy Trimester Tissues Trabecular meshwork structure Transcript Transforming Growth Factor beta Transgenic Mice Translational Research Validation Vinculin Vision Western Blotting Work aqueous causal variant cell behavior crosslink design differential expression early onset in vivo infancy innovation interest knock-down morphometry novel optic nerve disorder perinatal period postnatal preservation pressure prevent primary congenital glaucoma protein expression public health relevance targeted treatment transcriptomic profiling transcriptomics translational model translational potential treatment strategy vector control

项目摘要

Summary: A unique animal model of glaucoma with spontaneously arising mutation in LTBP2 has been established. This gene is implicated in heterogeneous glaucoma phenotypes, including human primary congenital glaucoma (PCG) as well as some forms of adult-onset open angle glaucoma. As the only known large animal homolog of inherited glaucoma in humans, this model has considerable potential for translational research. Posterior segment pathology has been extensively characterized and closely recapitulates features of human glaucomatous optic neuropathy. However, the role played by LTBP2 in the eye remains unknown and the precise cellular and molecular mechanisms responsible for the underlying elevated intraocular pressure (IOP) in this form of glaucoma remain unresolved. The objective of this proposal is to identify and localize, as potential targets for therapeutic intervention, key pathobiological mechanisms that contribute to both early onset and progressive IOP elevation in this model. In addition to perturbing LTBP2’s acknowledged role in microfibril assembly, enhanced TGF beta signaling, contributing to extracellular matrix (ECM) accumulation within the trabecular meshwork (TM) has been proposed as a mechanism for IOP elevation in this “microfibrillopathy”. However, preliminary studies in our animal model have not established whether significant increase in aqueous humor TGF beta concentrations represent a cause or effect of IOP elevation. Our central hypothesis is that LTBP2 is both an important regulator of outflow pathway development and remodeling in the perinatal period and a regulator of TM cell and ECM biology and pro-fibrotic pathways throughout life, extending beyond development. We will pursue two specific Aims: Aim 1: To determine the effects of absent or reduced LTBP2 expression on development of proximal and distal aqueous outflow pathways, we will test the hypothesis that LTBP2 plays an important role in outflow pathway development and remodeling using a combination of in vivo and in situ approaches to examine distal outflow morphology and ultrastructure of the proximal outflow pathway (TM and juxtacanalicular tissue) and ECM components of these pathways in wildtype and LTBP2+/- and LTBP2-/- eyes. Aim 2: To determine the pathological effects of absent or reduced LTBP2 expression on TM cell (TMC) biology, we will use TMC cultures derived from wildtype, LTBP2+/- and LTBP2-/- eyes. In Aim 2, we will test the hypotheses that reduced LTBP2 expression will a) negatively impact TMC attachment and proliferation in vitro and b) enhance the pathologic effects of TGFβ on actin cytoskeleton and promotion of ECM fibrosis. Successful completion of these Aims will generate new knowledge on the complex role of LTBP2 in physiology and pathology of the aqueous outflow pathways, providing a critical foundation to advance our overall goal of designing and testing innovative therapeutic approaches to glaucoma in our unique translational model.

概括：已经建立了一种具有 LTBP2 自发突变的独特青光眼动物模型。该基因与异质性青光眼表型有关，包括人类原发性先天性青光眼 (PCG) 以及某些形式的成人发病的开角型青光眼作为唯一已知的大型动物同源物。人类遗传性青光眼，该模型具有巨大的转化研究潜力。节段病理学已被初步表征并密切概括了人类的特征然而，LTBP2 在眼睛中所起的作用仍不清楚。导致眼内压 (IOP) 升高的精确细胞和分子机制这种形式的青光眼仍未得到解决，该提案的目标是识别和定位。治疗干预的潜在目标、有助于早期治疗的关键病理生物学机制除了扰乱 LTBP2 的公认作用外，该模型中的发病和渐进性 IOP 升高。微原纤维组装，增强 TGF β 信号传导，有助于细胞外基质 (ECM) 积累小梁网（TM）内的小梁网（TM）已被提议作为眼压升高的机制。然而，我们的动物模型的初步研究尚未确定是否显着。房水中 TGF β 浓度的增加代表了 IOP 升高的原因或影响。假设 LTBP2 是流出通路发育和重塑的重要调节因子围产期以及 TM 细胞和 ECM 生物学以及整个生命过程中促纤维化途径的调节器，我们将追求两个具体目标：目标 1：确定缺席的影响。或者近端和远端房水流出通路发育时 LTBP2 表达减少，我们将测试假设 LTBP2 在流出通路的发展和重塑中发挥重要作用结合体内和原位方法来检查远端流出形态和超微结构野生型中近端流出途径（TM 和近管组织）和这些途径的 ECM 成分目标 2：确定 LTBP2 缺失或减少的病理影响。 TM 细胞 (TMC) 生物学上的表达，我们将使用源自野生型、LTBP2+/- 和 LTBP2-/- 的 TMC 培养物在目标 2 中，我们将测试减少 LTBP2 表达将 a) 对 TMC 产生负面影响的假设。体外附着和增殖 b) 增强 TGFβ 对肌动蛋白细胞骨架的病理作用促进 ECM 纤维化的成功完成将产生关于复杂的新知识。 LTBP2 在房水流出途径的生理学和病理学中的作用，为我们的总体目标是以我们独特的方式设计和推进测试青光眼的创新治疗方法翻译模型。