Epithelial membrane protein-2 (EMP2) and experimental proliferative vitreoretinopathy (PVR).
Epithelial membrane protein-2 (EMP2) and experimental proliferative vitreoretinopathy (PVR).
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DOI：
10.3109/02713683.2011.561468
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发表时间：
2011-06
影响因子：
2
通讯作者：
Gordon LK
中科院分区：
文献类型：
--
作者：
Telander DG;Morales SA;Mareninov S;Forward K;Gordon LK
增殖性玻璃体视网膜病变（PVR）被认为部分是由于视网膜色素上皮（RPE）去分化，细胞在玻璃体腔迁移、形成膜以及在异常的伤口愈合过程中收缩所致。在体外胶原凝胶收缩实验中，上皮膜蛋白2（EMP2）通过激活RPE细胞系（ARPE - 19）中的粘着斑激酶（FAK）来控制收缩。本研究的目的是在体内模型中探究阻断或改变EMP2表达水平如何改变临床PVR。
利用ARPE - 19细胞系，通过稳定转染EMP2过表达构建体、EMP2核酶或单独的载体来调节EMP2水平。这些转染的细胞系被用于PVR兔模型。PVR的严重程度由两名不知情的观察者进行分类。还使用了一种EMP2阻断抗体来降低PVR模型中EMP2的功能。免疫组织化学用于评估体内EMP2的表达。
EMP2水平较低的转染细胞所引起的PVR严重程度明显低于野生型细胞诱导的PVR程度（p = 0.05）。此外，EMP2低表达的转染细胞比高表达EMP2的转染细胞呈现出PVR严重程度更低的强烈趋势（p = 0.06）。用特异性多克隆抗体阻断EMP2可显著降低PVR的严重程度（p = 0.02）。发现PVR膜的EMP2表达呈阳性。
这些体内研究支持EMP2表达与PVR严重程度之间存在直接相关性。这些结果证实了通过改变EMP2表达来控制RPE生物学的可能性，并为这种疾病提供了一个潜在的治疗靶点。
Proliferative vitreoretinopathy (PVR) is believed to result in part from de-differentiation of retinal pigment epithelium (RPE) with cellular migration in the vitreous cavity, membrane formation, and contraction in an aberrant wound-healing strategy. In an in vitro collagen-gel contraction assay, epithelial membrane protein 2 (EMP2) controls contraction through activation of focal adhesion kinase (FAK) in a RPE cell line (ARPE-19). The purpose of this study was to investigate how blocking or altering the level of EMP2 expression changed clinical PVR in an in vivo model. Using the ARPE-19 cell line, the levels of EMP2 modulated through stable transfections of an EMP2 overexpressing construct, EMP2 ribozyme, or vector alone. These transfected cell lines were used in a rabbit model of PVR. The severity of PVR was classified by two masked observers. An EMP2 blocking antibody was also used to decrease functional EMP2 in the PVR model. Immunohistochemistry was used to evaluate EMP2 expression in vivo. The transfectants with lower levels of EMP2 had significantly less PVR severity than the degree of PVR induced by wild-type cells (p = 0.05). Also, the transfectants with a low-level of EMP2 expression showed a strong trend of less PVR severity than the high-levels EMP2 transfectants (p = 0.06). Blocking EMP2 with a specific polyclonal antibody significantly decreased the level of PVR severity (p = 0.02). PVR membranes were found to be positive for EMP2 expression. These in vivo studies support a direct correlation between EMP2 expression and severity of PVR. These results validate the potential for controlling RPE biology through a change in EMP2 expression, and provide a potential therapeutic target for this disease.