Selective Blockade of Cytoskeletal Actin Remodeling Reduces Experimental Choroidal Neovascularization
Selective Blockade of Cytoskeletal Actin Remodeling Reduces Experimental Choroidal Neovascularization
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DOI：
10.1167/iovs.10-6351
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发表时间：
2011-04-01
影响因子：
4.4
通讯作者：
Chaqour, Brahim
中科院分区：
文献类型：
--
作者：
Caballero, Sergio;Yang, Ru;Chaqour, Brahim
目的.检测肽Ac-EEED在体外减少细胞粘附和增殖以及在体内减少脉络膜新生血管（CNV）的功效。将含有Ac-EEED序列的肽嵌合体化学连接至来自E-rns病毒表面蛋白的XMTM递送肽的N末端。将Ac-EEED或乱序对照肽（SCRAM）加入血管平滑肌细胞、周细胞、内皮细胞和成纤维细胞的培养物中，并评估粘附、生长和基质产生。将Ac-EEED或SCRAM注射到经历Bruch膜激光破裂的小鼠的玻璃体中以诱导CNV，并评估损伤体积、新生血管形成和损伤纤维化。Ac-EEED通过抑制G-肌动蛋白的聚合和破坏应力纤维的形成来诱导肌动蛋白细胞骨架形态的变化。用Ac-EEED预处理导致内皮细胞对VEGF的促有丝分裂和促粘附作用的反应降低。通过结缔组织生长因子、富含半胱氨酸的61、胶原蛋白I（COL 1A 2）和胶原蛋白III（COL 3A 1）的水平降低来评估，成纤维细胞中的Ac-EEED处理减少了TGF-β诱导的纤维化。CNV病变大小和纤维化以浓度依赖性方式减少高达60%。体外研究表明，Ac-EEED影响与细胞骨架肌动蛋白相关的广泛的机械性能，以减少生长因子的影响。Ac-EEED在体内的应用可能提供一种新的治疗策略，既抑制新生血管生长，又减少通常与CNV的退化阶段相关的纤维化。（Invest Ophthalmol维斯科学。2011; 52：2490-2496）DOI：10.1167/iovs.10-6351
PURPOSE. The efficacy of the peptide Ac-EEED on reducing cell adhesion and proliferation in vitro and choroidal neovascularization (CNV) in vivo was examined.METHODS. The peptide chimera containing the Ac-EEED sequence was chemically linked to the N terminus of the XMTM delivery peptide from the E-rns viral surface protein. Ac-EEED or scrambled control peptide (SCRAM) was added to cultures of vascular smooth muscle cells, pericytes, endothelial cells, and fibroblasts, and adhesion, growth, and matrix production was assessed. Ac-EEED or SCRAM was injected into the vitreous of mice undergoing laser rupture of Bruch's membrane to induce CNV and lesion volume, neovascularization and lesion fibrosis were assessed.RESULTS. Ac-EEED-induced changes in the morphology of the actin cytoskeleton by inhibiting polymerization of G-actin and disrupting the formation of stress fibers. Pretreatment with Ac-EEED resulted in endothelial cells becoming less responsive to the mitogenic and pro-adhesive effects of VEGF. Ac-EEED treatment in fibroblasts reduced TGF-beta-induced fibrosis as assessed by decreased levels of connective tissue growth factor, cysteine-rich 61, collagen I (COL1A2), and collagen III (COL3A1). CNV lesion size and fibrosis were reduced in a concentration-dependent manner by up to 60%.CONCLUSIONS. In vitro studies showed that Ac-EEED affects a broad range of mechanical properties associated with cytoskeletal actin to reduce growth factor effects. The utilization of Ac-EEED in vivo may offer a novel therapeutic strategy by both suppressed neovessel growth and curtailing fibrosis typically associated with the involutional stage of CNV. (Invest Ophthalmol Vis Sci. 2011; 52: 2490-2496) DOI: 10.1167/iovs.10-6351