Trabecular Meshwork Cytoskeletal Signaling-Regulation of Aqueous Humor Outflow

小梁网细胞骨架信号传导-房水流出的调节

• 批准号: 10337195
• 负责人: P VASANTHA Rao
• 金额: $ 39.04万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10337195
• 关键词: 2-tyrosine; Actins; Address; Adhesions; Adhesives; Antihypertensive Agents; Aqueous Humor; Atomic Force Microscopy; Atrophic; Biochemical; Biological; Biophysics; Blindness; Cataract; Cell Adhesion; Cell Shape; Cell surface; Cells; Cellular biology; Characteristics; Cytoskeletal Modeling; Deposition; Development; Dexamethasone; Disease; Disintegrins; Enzymes; Etiology; Evaluation; Extracellular Domain; Extracellular Matrix; Extracellular Matrix Proteins; Eye; Focal Adhesion Kinase 1; Funding; Gene Targeting; Glaucoma; Goals; Histologic; Homeostasis; Homologous Protein; Human; Impairment; Integrins; Intercellular Junctions; Investigation; Lead; Link; MADH2 gene; Mass Spectrum Analysis; Matrix Metalloproteinases; Mediating; Metalloproteases; Modification; Molecular; Ocular Hypertension; Optic Nerve; PTEN gene; PTK2 gene; Pathologic; Pathway interactions; Patients; Permeability; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiologic Intraocular Pressure; Physiological; Play; Post-Translational Protein Processing; Production; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Proteoglycan; Proteomics; Rattus; Regulation; Resistance; Rho-associated kinase; Risk Factors; Role; Sampling; Secretory Cell; Signal Pathway; Signal Transduction; Time; Trabecular meshwork structure; Transforming Growth Factor Beta 2; Tyrosine Phosphorylation; base; cell behavior; cell type; efficacious treatment; insight; microscopic imaging; novel; novel strategies; ocular hypotensive; response; rho; sight restoration; syndecan; treatment strategy

PROJECT SUMMARY Dysregulated homeostatic mechanisms that lead to an increased resistance to aqueous humor (AH) outflow through the trabecular pathway constitute the main cause for ocular hypertension (OHT), a key risk factor for an irreversible blinding disease, glaucoma. The extracellular matrix (ECM) is well-recognized to play a crucial role in modulation of AH outflow and intraocular pressure (IOP) via regulating the contractile, adhesive, fibrogenic and permeability characteristics of the trabecular outflow pathway cells. There still exists a fundamental gap in our understanding however, of how post-translational modifications of ECM proteins and ECM degrading enzymes might influence trabecular meshwork (TM) cell behavior, AH outflow and IOP. The broad objective of this proposal is to examine protein phosphorylation of ECM components and matrix metalloproteinases (MMPs), and determine how this modification modulates AH outflow and IOP, with the goal of uncovering novel insights into OHT etiology and developing efficacious strategies for glaucoma treatment. The scientific premise is that protein tyrosine kinases and phosphatases in the AH secreted by TM and other cell types play a vital role in homeostasis of AH outflow and IOP by modifying protein tyr-phosphorylation status of ECM and MMPs that in turn influences TM cell contractile, adhesive, fibrogenic and barrier activities. We hypothesize that deregulation of tyr-phosphorylation of ECM proteins and MMPs resulting from alterations in the levels of secretory tyrosine kinases and phosphatases may be a key etiological contribution to impaired AH outflow and elevated IOP in glaucoma. In support of this novel and paradigm shifting hypothesis, our recent studies detected vertebrate lonesome kinase (VLK) and phosphatase and tensin homolog (PTEN) proteins in human AH samples, and demonstrated that HTM cells secrete both enzymes. Significantly, VLK was shown to regulate ECM tyr-phosphorylation, and contractile and adhesive activities of TM cells, and induced by TGF-β2 and dexamethasone in human TM cells, both of which are known to induce OHT. Based on these promising and novel preliminary results, we have outlined three specific aims in this competing proposal, focused on mechanistic investigations into the role(s) of VLK in: 1) regulation of intracellular signaling pathways controlling TM cell contractile, cell-ECM & cell-cell adhesive, fibrogenic and permeability characteristics, 2) Tyr-phosphorylation of TM cell ECM and MMPs, ECM organization and TM cell stiffness, and 3) AH outflow and IOP, using human TM cells, human donor eyes and rats. These studies will utilize gene targeting, proteomics, biophysical, histological and physiological approaches, and include an assessment of changes in levels of VLK and PTEN in AH samples from glaucoma versus non-glaucoma (cataract) patients. Completion of the novel studies proposed in this application is expected to uncover new understanding on the role of tyr-phosphorylation of ECM proteins and MMPs in AH outflow and OHT etiology via altered outside-in signaling, and to identify new and efficacious treatment strategies for lowering IOP in glaucoma patients.

项目概要 体内平衡机制失调导致房水 (AH) 流出阻力增加 通过小梁通路是高眼压症（OHT）的主要原因，而高眼压症是高眼压症的一个关键危险因素。 众所周知，青光眼是一种不可逆的致盲疾病。 通过调节收缩、粘着、 小梁流出途径细胞的纤维形成和通透性特征仍然存在。 然而，我们对 ECM 蛋白的翻译后修饰如何 ECM 降解酶可能影响小梁网 (TM) 细胞行为、AH 流出和 IOP。 该提案的主要目标是检查 ECM 成分和基质的蛋白质磷酸化 金属蛋白酶 (MMP)，并确定这种修饰如何调节 AH 流出和 IOP，目标是 揭示 OHT 病因的新见解并制定青光眼治疗的有效策略。 科学前提是TM等分泌的AH中的蛋白酪氨酸激酶和磷酸酶 细胞类型通过改变蛋白质酪氨酸磷酸化在 AH 流出和 IOP 的稳态中发挥重要作用 ECM 和 MMP 的状态反过来影响 TM 细胞的收缩、粘附、纤维形成和屏障活性。 我们追求因改变而导致 ECM 蛋白和 MMP 酪氨酸磷酸化的失调 分泌性酪氨酸激酶和磷酸酶水平可能是导致受损的关键病因 为了支持这一新颖且范式转变的假设，我们最近研究了青光眼的 AH 流出和眼压升高。 研究检测到脊椎动物孤独激酶（VLK）以及磷酸酶和张力蛋白同源物（PTEN）蛋白 人类 AH 样本，并证明 HTM 细胞分泌这两种酶。 调节 ECM 酪氨酸磷酸化以及 TM 细胞的收缩和粘附活性，并由 TGF-β2 诱导 人类 TM 细胞中的地塞米松和地塞米松，已知这两种物质都会诱导 OHT。 基于这些有希望且新颖的初步结果，我们在本次竞争中概述了三个具体目标 提案，重点是 VLK 在以下方面的作用的机制研究：1) 细胞内信号传导的调节 控制途径 TM 细胞收缩、细胞-ECM 和细胞-细胞粘附、纤维形成和渗透性 特征，2) TM 细胞 ECM 和 MMP 的酪氨酸磷酸化、ECM 组织和 TM 细胞硬度， 3) 使用人类 TM 细胞、人类供体眼睛和大鼠进行 AH 流出和 IOP 这些研究将利用基因。 靶向、蛋白质组学、生物物理、组织学和生理学方法，并包括评估 青光眼患者与非青光眼（白内障）患者的 AH 样本中 VLK 和 PTEN 水平的变化。 本申请中提出的新颖研究的完成预计将揭示对 ECM 蛋白和 MMP 酪氨酸磷酸化通过由外向内改变在 AH 流出和 OHT 病因学中的作用 信号传导，并确定降低青光眼患者眼压的新的有效治疗策略。

项目成果

Elevated intraocular pressure induces Rho GTPase mediated contractile signaling in the trabecular meshwork.

眼压升高会诱导小梁网中 Rho GTPase 介导的收缩信号传导。

• 发表时间: 2015-07
• 影响因子: 3.4
• 作者: Pattabiraman, Padmanabhan P;Inoue, Toshihiro;Rao, P Vasantha
• 通讯作者: Rao, P Vasantha

Role of MCP-1 and IL-8 in viral anterior uveitis, and contractility and fibrogenic activity of trabecular meshwork cells.

MCP-1 和 IL-8 在病毒性前葡萄膜炎中的作用以及小梁网细胞的收缩性和纤维形成活性。

• 发表时间: 2021
• 影响因子: 4.6
• 作者: Lee, Jiyoung;Choi, Jin A;Ju, Hyun;Kim, Ju;Paik, Soon;Rao, Ponugoti Vasantha
• 通讯作者: Rao, Ponugoti Vasantha

Global phosphotyrosinylated protein profile of cell-matrix adhesion complexes of trabecular meshwork cells.

小梁网细胞的细胞基质粘附复合物的整体磷酸酪氨酸化蛋白质谱。

• 发表时间: 2020
• 作者: Maddala, Rupalatha;Rao, Ponugoti Vasantha
• 通讯作者: Rao, Ponugoti Vasantha

Influence of Growth Differentiation Factor 15 on Intraocular Pressure in Mice.

生长分化因子 15 对小鼠眼压的影响。

• 发表时间: 2024-04
• 作者: Maddala, Rupalatha;Eldawy, Camelia;Ho, Leona T Y;Challa, Pratap;Rao, Ponugoti V
• 通讯作者: Rao, Ponugoti V

Discovery of substituted 4-(pyrazol-4-yl)-phenylbenzodioxane-2-carboxamides as potent and highly selective Rho kinase (ROCK-II) inhibitors.

发现取代的 4-(吡唑-4-基)-苯基苯并二恶烷-2-甲酰胺作为有效且高选择性的 Rho 激酶 (ROCK-II) 抑制剂。

• DOI: 10.1021/jm800986w
• 发表时间: 2008-11-13
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Feng Y;Yin Y;Weiser A;Griffin E;Cameron MD;Lin L;Ruiz C;Schürer SC;Inoue T;Rao PV;Schröter T;Lograsso P
• 通讯作者: Lograsso P