Role of Matrix Metalloproteinases in Subcapsular Cataract Formation

基质金属蛋白酶在囊下白内障形成中的作用

基本信息

批准号：
7825296
负责人：
Judith A West-Mays
金额：
$ 20.76万
依托单位：
MCMASTER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7825296
关键词：
Anterior Cataract Conditioned Culture Media Crystalline Lens Data Development Disease E-Cadherin Epithelial Epithelial Cells Exhibits Family member Fibrosis Figs - dietary Gelatinase A Gelatinase B Genes Genetic Goals In Vitro Knockout Mice MAP Kinase Gene MAP Kinase Signaling Pathways MAPK14 gene MMPI Malignant Neoplasms Matrix Metalloproteinases Mediating Mesenchymal Mitogen-Activated Protein Kinases Modeling Mus Mutant Strains Mice Myofibroblast Pathology Pathway interactions Phenotype Phosphorylation Play Rattus Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signal Transduction Signaling Molecule Smooth Muscle Actin Staining Method System Testing Time Western Blotting Wound Healing cytokine in vitro Model in vivo kinase inhibitor laser capture microdissection lens morphogens mouse model prevent programs repaired

项目摘要

The cytokine TGFP is a pleotropic morphogen that modulates the tissue repair phenotype and also plays an important role in the development of fibrotic repair pathologies. In the lens of the eye, fibrotic pathologies mediated by TGFP include anterior subcapsular cataracts (ASC) and posterior capsular opacification (PCO). The cellular changes that precede fibrosis in ASC and PCO include an increased proliferation of lens epithelial cells (LECs), which under go an epithelial-mesenchymal transformation (EMT) into myofibroblasts, involving loss of E-cadherin and induced a-smooth muscle actin (aSMA) expression. The long-term goal of this project is to determine the TGFp-mediated signals, which alter the genetic makeup and phenotype of lens epithelial cells during ASC. Using a previously developed rat lens culture model in which exogenous TGFP induces ASC we have shown that treatment with agents that inhibit enzymatic activity of Matrix Metalloproteinase (MMP) family members (MMPIs) suppresses the TGFp-induced cataractous changes. Further preliminary findings suggest the testable hypothesis that MMPIs act to inhibit TGFP- induced EMT and ASC in the lens by suppressing MMP-mediated E-cadherin degradation by shedding. The RSmad, SmadS, is a common effector of TGFP signaling, which has been identified in the lens. However, we have found using two different mouse models that in the absence of SmadS (in Smad3 KO mice) TGFpl can induce EMT in the lens, demonstrating the involvement of SmadS-independent signaling in ASC formation. Additional findings suggest that the hypothesis that TGFp-induced ASC involves signaling MAP kinase pathways, specifically p38,which act independently of Smad3. To test these hypotheses we will utilize the in vitro TGFp-induced rat lens model of ASC, as well as, genetically modified mice, including MMP-2, MMP-9 and SmadS KO mice. The effect of MMPIs on E-cadherin expression and shedding will be further examined using RT-QPCR in combination with laser capture microdissection, western blotting and immunolocalization. The requirement for activated pSSMAPKin ASC formation will be investigated in both the in vivo and in vitro models using specific Pp38 kinase inhibitors. These data will aid in defining the TGFp-mediated pathways controlling EMT and fibrosis in ASC. Furthermore, since the genes and signaling mechanisms in ASC formation are similar to those, which occur, in other fibrotic diseases and cancer, the information gained from these studies in the lens, may also have relevance to these disease entities.

细胞因子 TGFP 是一种多效性形态发生素，可调节组织修复表型，并发挥重要作用。在纤维化修复病理学的发展中发挥重要作用。在眼睛的晶状体中，纤维化病变 TGFP介导的白内障包括前囊膜下白内障（ASC）和后囊膜混浊（PCO）。 ASC 和 PCO 纤维化之前的细胞变化包括晶状体增殖增加上皮细胞（LEC），经过上皮间质转化（EMT）肌成纤维细胞，涉及 E-钙粘蛋白的丢失和诱导的 a-平滑肌肌动蛋白 (aSMA) 表达。这该项目的长期目标是确定 TGFp 介导的信号，这些信号会改变基因组成和 ASC 期间晶状体上皮细胞的表型。使用先前开发的大鼠晶状体培养模型，其中外源性 TGFP 诱导 ASC 我们已经证明，用抑制酶活性的药物进行治疗基质金属蛋白酶 (MMP) 家族成员 (MMPI) 抑制 TGFp 诱导的白内障变化。进一步的初步研究结果提出了可检验的假设，即 MMPI 可以抑制 TGF-β 通过抑制 MMP 介导的 E-钙粘蛋白降解诱导晶状体中的 EMT 和 ASC 脱落。 RSmad、SmadS 是 TGFP 信号传导的常见效应器，已在镜片。然而，我们发现使用两种不同的小鼠模型，在缺乏 SmadS 的情况下（在 Smad3 KO 小鼠）TGFpl可以诱导晶状体中的EMT，证明SmadS独立信号传导参与 ASC 形成。其他研究结果表明，TGFβ 诱导的 ASC 涉及信号传导的假设 MAP 激酶途径，特别是 p38，其作用独立于 Smad3。为了检验这些假设，我们将利用体外 TGFβ 诱导的 ASC 大鼠晶状体模型以及转基因小鼠，包括 MMP-2、MMP-9 和 SmadS KO 小鼠。 MMPI 对 E-cadherin 表达和脱落的影响使用 RT-QPCR 结合激光捕获显微切割、蛋白质印迹和免疫定位。将在以下两个方面研究激活 pSSMAPKin ASC 形成的要求使用特定 Pp38 激酶抑制剂的体内和体外模型。这些数据将有助于定义 TGFβ 介导的途径控制 ASC 中的 EMT 和纤维化。此外，由于基因和信号传导 ASC 形成的机制与其他纤维化疾病和癌症中发生的机制相似，从这些镜头研究中获得的信息，也可能与这些疾病实体相关。