Peiris-Pagès Maria, Ozsvári Béla, Sotgia Federica, Lisanti Michael P
Clinical and Experimental Pharmacology, University of Manchester, Cancer Research UK, Manchester, United Kingdom.
Translational Medicine, School of Environment and Life Sciences, Biomedical Research Centre (BRC), University of Salford, Greater Manchester, United Kingdom.
Aging (Albany NY). 2019 Jul 16;11(14):4801-4835. doi: 10.18632/aging.102054.
Using proteomics analysis, we previously compared MCF7 breast cancer cells grown as 3D tumor spheres, with the same cell line grown as monolayers. Our results indicated that during 3D anchorage-independent growth, the cellular machinery associated with i) mitochondrial biogenesis and ii) ribosomal biogenesis, were both significantly increased. Here, for simplicity, we refer to these two new oncogenic hallmarks as "mito-stemness" and "ribo-stemness" features. We have now applied this same type of strategy to begin to understand how fibroblasts and MCF7 breast cancer cells change their molecular phenotype, when they are co-cultured together. We have previously shown that MCF7-fibroblast co-cultures are a valuable model of resistance to apoptosis induced by hormonal therapies, such as Tamoxifen and Fulvestrant. Here, we directly show that these mixed co-cultures demonstrate the induction of mito-stemness and ribo-stemness features, likely reflecting a mechanism for cancer cells to increase their capacity for accumulating biomass. In accordance with the onset of a stem-like phenotype, KRT19 (keratin 19) was induced by ~6-fold during co-culture. KRT19 is a well-established epithelial CSC marker that is used clinically to identify metastatic breast cancer cells in sentinel lymph node biopsies. The potential molecular therapeutic targets that we identified by label-free proteomics of MCF7-fibroblast co-cultures were then independently validated using a bioinformatics approach. More specifically, we employed publically-available transcriptional profiling data derived from primary tumor samples from breast cancer patients, which were previously subjected to laser-capture micro-dissection, to physically separate breast cancer cells from adjacent tumor stroma. This allowed us to directly validate that the proteins up-regulated in this co-culture model were also transcriptionally elevated in patient-derived breast cancer cells . This powerful approach for target identification and translational validation, including the use of patient outcome data, can now be applied to other tumor types as well, to validate new therapeutic targets that are more clinically relevant, for patient benefit. Moreover, we discuss the therapeutic implications of these findings for new drug development, drug repurposing and Tamoxifen-resistance, to effectively target mito-stemness and ribo-stemness features in breast cancer patients. We also discuss the broad implications of this "organelle biogenesis" approach to cancer therapy.
我们之前通过蛋白质组学分析，比较了以三维肿瘤球形式生长的MCF7乳腺癌细胞与以单层形式生长的同细胞系。我们的结果表明，在三维非锚定依赖性生长过程中，与i）线粒体生物发生和ii）核糖体生物发生相关的细胞机制均显著增加。在此，为简便起见，我们将这两个新的致癌标志称为“线粒体干性”和“核糖体干性”特征。我们现在应用了相同类型的策略，以开始了解成纤维细胞和MCF7乳腺癌细胞在共同培养时如何改变其分子表型。我们之前已经表明，MCF7-成纤维细胞共培养是对他莫昔芬和氟维司群等激素疗法诱导的凋亡产生抗性的有价值模型。在此，我们直接表明，这些混合共培养物显示出线粒体干性和核糖体干性特征的诱导，这可能反映了癌细胞增加其积累生物量能力的一种机制。与干细胞样表型的出现一致，在共培养期间KRT19（角蛋白19）被诱导增加了约6倍。KRT19是一种成熟的上皮性癌症干细胞标志物，临床上用于在前哨淋巴结活检中识别转移性乳腺癌细胞。然后，我们使用生物信息学方法独立验证了通过MCF7-成纤维细胞共培养的无标记蛋白质组学鉴定出的潜在分子治疗靶点。更具体地说，我们采用了来自乳腺癌患者原发性肿瘤样本的公开转录谱数据，这些样本之前经过激光捕获显微切割，以便从相邻肿瘤基质中物理分离乳腺癌细胞。这使我们能够直接验证在这种共培养模型中上调的蛋白质在患者来源的乳腺癌细胞中也在转录水平上升高。这种强大的靶点识别和转化验证方法，包括使用患者预后数据，现在也可以应用于其他肿瘤类型，以验证更具临床相关性的新治疗靶点，从而使患者受益。此外，我们讨论了这些发现对新药开发、药物再利用和他莫昔芬耐药性的治疗意义，以有效靶向乳腺癌患者中的线粒体干性和核糖体干性特征。我们还讨论了这种“细胞器生物发生”方法对癌症治疗的广泛影响。
