Screening assay for blood vessel maturation inhibitors.
Screening assay for blood vessel maturation inhibitors.
复制标题
血管成熟抑制剂的筛选测定。
DOI：
10.1016/j.bbrc.2013.07.077
复制
发表时间：
2013
影响因子：
3.1
通讯作者：
Hammers,HansJ
中科院分区：
文献类型：
--
作者：
Fu,Chenglai;vanderZwan,Anita;Gerber,Stephanie;VanDenBerg,Susan;No,Elisa;Wang,WayneCH;Sheibani,Nader;Carducci,MichaelA;Kachhap,Sushant;Hammers,HansJ
在癌症患者中，对靶向VEGF途径的抗血管生成剂的耐药性的发展是常见的。已经表明，接受VEGF靶向治疗的肿瘤脉管系统的周细胞覆盖增加在抵抗中起重要作用。因此，减少肿瘤脉管系统的周细胞覆盖已被认为是打破抗血管生成疗法的抗性和增加抗血管生成疗法的功效的治疗方法。为了筛选化合物库，需要血管成熟的简单体外测定，其证明内皮细胞和周细胞缔合同时形成管腔化血管结构。不幸的是，从图像和数据采集的角度来看，先前描述的基于3维基质的测定是费力且具有挑战性的。由于这些原因，它们通常缺乏在高吞吐量环境中执行所需的可伸缩性。通过这项工作，我们开发了一种新的体外血管成熟试验，其中管腔化的血管结构在一个光学平面上形成，间充质祖细胞（10 T1/2）分化为周细胞样细胞，其与内皮血管（HUVECs）相关联。使用由α平滑肌肌动蛋白启动子（SMP-8）控制的GFP报告子，观察10 T1/2细胞分化成周细胞样细胞。在一个光学平面中组织这些血管结构及其募集的壁细胞允许自动化数据捕获和随后的图像分析。验证了该试验筛选周细胞募集抑制剂的能力。总之，这种体外血管成熟的新方法为筛选具有治疗潜力的新药物提供了一种有价值的工具。
In cancer patients, the development of resistance to anti-angiogenic agents targeting the VEGF pathway is common. Increased pericyte coverage of the tumor vasculature undergoing VEGF targeted therapy has been suggested to play an important role in resistance. Therefore, reducing the pericytes coverage of the tumor vasculature has been suggested to be a therapeutic approach in breaking the resistance to and increasing the efficacy of anti-angiogenic therapies. To screen compound libraries, a simplein vitroassay of blood vessel maturation demonstrating endothelial cells and pericytes association while forming lumenized vascular structures is needed. Unfortunately, previously described 3-dimensional, matrix based assays are laborious and challenging from an image and data acquisition perspective. For these reasons they generally lack the scalability needed to perform in a high-throughput environment. With this work, we have developed a novelin vitroblood vessel maturation assay, in which lumenized, vascular structures form in one optical plane and mesenchymal progenitor cells (10T1/2) differentiate into pericyte-like cells, which associate with the endothelial vessels (HUVECs). The differentiation of the 10T1/2 cells into pericyte-like cells is visualized using a GFP reporter controlled by the alpha smooth muscle actin promoter (SMP-8). The organization of these vascular structures and their recruited mural cells in one optical plane allows for automated data capture and subsequent image analysis. The ability of this assay to screen for inhibitors of pericytes recruitment was validated. In summary, this novel assay ofin vitroblood vessel maturation provides a valuable tool to screen for new agents with therapeutic potential.