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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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要导致房水（AH）流出阻力增加的体内平衡机制失调通过小梁途径的视网膜病变构成了高眼压（OHT）的主要原因，一种不可逆转的致盲性疾病青光眼细胞外基质（ECM）是公认的发挥关键作用，在调节AH流出和眼内压（IOP）中的作用，通过调节收缩，粘附，小梁流出道细胞的纤维化和渗透性特征。仍存在然而，我们对ECM蛋白质的翻译后修饰以及ECM蛋白质的翻译后修饰的理解存在根本性的差距。 ECM降解酶可能影响小梁网细胞行为、AH流出和IOP。的该建议的广泛目标是检查ECM组分和基质的蛋白磷酸化金属蛋白酶（MMPs），并确定这种修饰如何调节AH流出和IOP，目标是发现OHT病因学的新见解，并制定有效的青光眼治疗策略。科学的前提是，TM和其他细胞分泌的AH中的蛋白酪氨酸激酶和磷酸酶，细胞类型通过修饰蛋白酪氨酸磷酸化在AH流出和IOP的稳态中起重要作用 ECM和MMPs的状态进而影响TM细胞收缩、粘附、纤维化和屏障活性。我们假设，由于细胞外基质蛋白和基质金属蛋白酶的改变，分泌性酪氨酸激酶和磷酸酶水平的下降可能是导致受损的关键病因。青光眼中的AH流出和IOP升高。为了支持这一新的和范式转变的假设，我们最近的研究检测了脊椎动物孤体激酶（VLK）和磷酸酶和张力蛋白同源物（PTEN）蛋白，人AH样品，并证明HTM细胞分泌这两种酶。值得注意的是，VLK被证明调节ECM酪氨酸磷酸化，调节TM细胞的收缩和粘附活性，并由TGF-β2诱导和地塞米松，已知这两种物质都能诱导OHT。基于这些有希望的和新的初步结果，我们概述了这一竞争的三个具体目标，该提案集中于对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TM细胞收缩、细胞-ECM和细胞-细胞粘附、纤维化和渗透性的途径 2）TM细胞ECM和MMP的Tyr-磷酸化，ECM组织和TM细胞硬度，和3）AH流出和IOP，使用人TM细胞、人供体眼和大鼠。这些研究将利用基因靶向、蛋白质组学、生物物理学、组织学和生理学方法，并包括评估来自青光眼与非青光眼（白内障）患者的AH样品中VLK和PTEN水平的变化。本申请中提出的新研究的完成有望揭示对 ECM蛋白和MMPs的酪氨酸磷酸化在AH流出和OHT病因学中通过由外向内改变的作用信号转导，并确定新的和有效的治疗策略，降低青光眼患者的IOP。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

DOI：
10.1038/s41598-021-94391-2
发表时间：
2021-07-22
期刊：
Scientific reports
影响因子：
4.6
作者：
Lee J;Choi JA;Ju HH;Kim JE;Paik SY;Rao PV
通讯作者：
Rao PV

The pulling, pushing and fusing of lens fibers: a role for Rho GTPases.

晶状体纤维的拉、推和融合：Rho GTPases 的作用。

DOI：
10.4161/cam.2.3.6495
发表时间：
2008
期刊：
Cell adhesion & migration
影响因子：
3.2
作者：
Rao,PVasantha
通讯作者：
Rao,PVasantha

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

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

Role of the Rho GTPase/Rho kinase signaling pathway in pathogenesis and treatment of glaucoma: Bench to bedside research.