Wenzel Carsten, Otto Saskia, Prechtl Stefan, Parczyk Karsten, Steigemann Patrick
Bayer Pharma AG, Global Drug Discovery, Berlin, Germany.
Bayer Pharma AG, Global Drug Discovery, Berlin, Germany.
Exp Cell Res. 2015 Nov 15;339(1):35-43. doi: 10.1016/j.yexcr.2015.10.003. Epub 2015 Oct 21.
Invasion processes underlie or accompany several pathological processes but only a limited number of high-throughput capable phenotypic models exist to test anti-invasive compounds in vitro. We here evaluated 3D co-cultures as a high-content phenotypic screening system for fibrotic invasive processes. 3D multicellular spheroids were used as living tissue surrogates in co-culture with fluorescently labeled lung fibroblasts to monitor invasion processes by automated microscopy. This setup was used to screen a compound library containing 480 known bioactive substances. Identified hits prevented fibroblast invasion and could be subdivided into two hit classes. First, Prostaglandins were shown to prevent fibroblast invasion, most likely mediated by the prostaglandin EP2 receptor and generation of cAMP. Additionally, Rho-associated protein kinase (ROCK) inhibitors prevented fibroblast invasion, possibly by inactivation of myosin II. Importantly, both Prostaglandins and ROCK inhibitors are potential treatment options shown to be effective in in vitro and in vivo models of fibrotic diseases. This validates the presented novel phenotypic screening approach for the evaluation of potential inhibitors and the identification of novel compounds with activity in diseases that are associated with fibroblast invasion.
侵袭过程是多种病理过程的基础或伴随现象，但目前仅有有限的高通量表型模型可用于体外测试抗侵袭化合物。我们在此评估了三维共培养作为纤维化侵袭过程的高内涵表型筛选系统。三维多细胞球体被用作与荧光标记的肺成纤维细胞共培养的活组织替代物，通过自动显微镜监测侵袭过程。该设置用于筛选包含480种已知生物活性物质的化合物库。鉴定出的活性物质可阻止成纤维细胞侵袭，并可分为两类。首先，前列腺素被证明可阻止成纤维细胞侵袭，最有可能是由前列腺素EP2受体和cAMP的生成介导的。此外，Rho相关蛋白激酶（ROCK）抑制剂可阻止成纤维细胞侵袭，可能是通过使肌球蛋白II失活。重要的是，前列腺素和ROCK抑制剂都是在纤维化疾病的体外和体内模型中显示有效的潜在治疗选择。这验证了所提出的新型表型筛选方法在评估潜在抑制剂和鉴定在与成纤维细胞侵袭相关疾病中具有活性的新型化合物方面的有效性。
