Control of Trabecular Meshwork Cytoskeleton

小梁网细胞骨架的控制

• 批准号: 7489937
• 负责人: Donna M Peters
• 金额: $ 34.97万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7489937
• 关键词: 1-Phosphatidylinositol 3-Kinase; Actins; Affect; Agonist; Antibodies; Aqueous Humor; Blood; Cells; Cultured Cells; Cytoskeleton; Data; Dominant-Negative Mutation; Ethacrynic Acid; Eye; Fibronectins; Glaucoma; Grant; Human; Inorganic Sulfates; Integrin Signaling Pathway; Integrins; Knowledge; Laboratories; Lead; Length; Ligands; Mediating; Modeling; Monkeys; Northern Blotting; Numbers; Organ Culture Techniques; Pathway interactions; Peptides; Phosphatidylinositol 4,5-Diphosphate; Physiologic Intraocular Pressure; Physiological; Play; Primary Open Angle Glaucoma; Protein Overexpression; Proteins; Proteoglycan; Regulation; Research Personnel; Retinal Ganglion Cells; Role; Signal Pathway; Signaling Molecule; Small Interfering RNA; Steroids; Stream; Stress Fibers; Thrombospondin 1; Trabecular meshwork structure; Unspecified or Sulfate Ion Sulfates; Up-Regulation; Vinblastine; crosslink; cytochalasin H; extracellular; inhibiting antibody; novel; optic nerve disorder; prevent; rac1 GTP-Binding Protein; receptor; syndecan; syndecan-4

DESCRIPTION: Glaucoma is an optic neuropathy that affects nearly 67 million people world-wide. It is commonly associated with elevated levels of intraocular pressure due to a reduction in aqueous humor outflow from the trabecular meshwork (TM). Recent studies indicate that the actin cytoskeleton has a profound influence on aqueous humor outflow and thus might provide a new target for the treatment for glaucoma. Integrin/syndecan mediated signaling pathways play a critical role in regulating the actin cytoskeleton. We propose that integrins and/or syndecans contribute to the regulation of the actin cytoskeleton and outflow facility and that these receptors may be responsible for changes in the cytoskeleton (i.e., CLAN formation) observed after steroid treatment. We further propose that steroid treatment activates these signaling pathways because it causes an upregulation of extracellular ligands or cytoplasmic signaling molecules associated with integrins and syndecans. The main objectives of this grant are to determine the role that integrins and syndecans play in governing the organization of the actin cytoskeleton in the TM and the extent to which they can be manipulated to regulate outflow facility. The aims of this grant are (1) determine the specific integrins responsible for the formation of CLANs in TM by using integrin specific peptides (agonists and antagonists) and activating or inhibiting antibodies to block or trigger CLAN formation, (2) use integrin antagonists and agonists to demonstrate that specific integrin signaling pathways influence outflow facility, especially in steroid treated eyes, (3) demonstrate that syndecan-4 is a key regulator of actin networks in the TM by blocking or enhancing its normal function using syndecan-4 siRNA and overexpression of full length or truncated syndecan-4 in TM cell cultures, (4) determine if virally expressed hairpin siRNA sequences to syndecan-4 would block steroid induced clan formation and decreases in outflow facility, and (5) determine if steroid treatment affects the expression or activity of proteins (Rac1, Tiam1, Trio, PI-3 kinase, Src, PIP2) associated with these pathways. Ultimately, this knowledge can be used to develop new therapies to treat or prevent POAG or steroid-induced glaucoma.

描述：青光眼是一种神经病，在全球范围内影响近6700万人。它通常与小梁网（TM）水性幽默流出降低引起的眼内压水平有关。最近的研究表明，肌动蛋白细胞骨架对水性幽默流出有深远的影响，因此可能为青光眼治疗提供了新的靶标。整联蛋白/syndecan介导的信号通路在调节肌动蛋白细胞骨架中起关键作用。我们建议整联蛋白和/或联合群有助于调节肌动蛋白细胞骨架和流出设施，并且这些受体可能导致固醇治疗后观察到的细胞骨架（即氏族形成）的变化。我们进一步提出类固醇治疗会激活这些信号通路，因为它会导致与整合素和联合素相关的细胞外配体或细胞质信号分子的上调。 这笔赠款的主要目的是确定整联蛋白和联合体在TM中肌动蛋白细胞骨架的组织中扮演的角色以及可以操纵它们以调节流出设施的程度。这笔赠款的目的是（1）通过使用整合素特定的肽（激动剂和拮抗剂）来确定负责TM中氏族形成的特定整合素TM中肌动蛋白网络的调节因子通过使用Syndecan-4 siRNA阻断或增强其正常功能，并过表达TM细胞培养中的全长或截短syndecan-4，（4）（4）病毒表达的发夹siRNA序列确定syndecan-4是否会阻止类固醇的固定剂的表现，并确定固定剂的表现，并确定（5）的表现（5） （Rac1，Tiam1，Trio，Pi-3激酶，SRC，PIP2）与这些途径相关联。最终，这些知识可用于开发新的疗法，以治疗或预防POAG或类固醇引起的青光眼。