Schwartz Alyssa D, Barney Lauren E, Jansen Lauren E, Nguyen Thuy V, Hall Christopher L, Meyer Aaron S, Peyton Shelly R
Department of Chemical Engineering, University of Massachusetts Amherst, 686 N Pleasant St. 159 Goessmann Laboratory, Amherst, MA 01003, USA.
Integr Biol (Camb). 2017 Dec 11;9(12):912-924. doi: 10.1039/c7ib00128b.
Traditional drug screening methods lack features of the tumor microenvironment that contribute to resistance. Most studies examine cell response in a single biomaterial platform in depth, leaving a gap in understanding how extracellular signals such as stiffness, dimensionality, and cell-cell contacts act independently or are integrated within a cell to affect either drug sensitivity or resistance. This is critically important, as adaptive resistance is mediated, at least in part, by the extracellular matrix (ECM) of the tumor microenvironment. We developed an approach to screen drug responses in cells cultured on 2D and in 3D biomaterial environments to explore how key features of ECM mediate drug response. This approach uncovered that cells on 2D hydrogels and spheroids encapsulated in 3D hydrogels were less responsive to receptor tyrosine kinase (RTK)-targeting drugs sorafenib and lapatinib, but not cytotoxic drugs, compared to single cells in hydrogels and cells on plastic. We found that transcriptomic differences between these in vitro models and tumor xenografts did not reveal mechanisms of ECM-mediated resistance to sorafenib. However, a systems biology analysis of phospho-kinome data uncovered that variation in MEK phosphorylation was associated with RTK-targeted drug resistance. Using sorafenib as a model drug, we found that co-administration with a MEK inhibitor decreased ECM-mediated resistance in vitro and reduced in vivo tumor burden compared to sorafenib alone. In sum, we provide a novel strategy for identifying and overcoming ECM-mediated resistance mechanisms by performing drug screening, phospho-kinome analysis, and systems biology across multiple biomaterial environments.
传统的药物筛选方法缺乏有助于产生耐药性的肿瘤微环境特征。大多数研究深入考察单一生物材料平台中的细胞反应，却在理解诸如硬度、维度和细胞间接触等细胞外信号如何独立作用或在细胞内整合以影响药物敏感性或耐药性方面存在空白。这至关重要，因为适应性耐药至少部分是由肿瘤微环境的细胞外基质（ECM）介导的。我们开发了一种方法来筛选在二维和三维生物材料环境中培养的细胞的药物反应，以探索ECM的关键特征如何介导药物反应。该方法发现，与水凝胶中的单细胞和塑料上的细胞相比，二维水凝胶上的细胞以及封装在三维水凝胶中的球体对靶向受体酪氨酸激酶（RTK）的药物索拉非尼和拉帕替尼反应较小，但对细胞毒性药物反应并非如此。我们发现，这些体外模型与肿瘤异种移植之间的转录组差异并未揭示ECM介导的对索拉非尼耐药的机制。然而，对磷酸化激酶组数据的系统生物学分析发现，MEK磷酸化的变化与RTK靶向药物耐药性相关。以索拉非尼作为模型药物，我们发现与MEK抑制剂联合给药在体外降低了ECM介导的耐药性，并且与单独使用索拉非尼相比，减轻了体内肿瘤负担。总之，我们提供了一种新策略，通过在多个生物材料环境中进行药物筛选、磷酸化激酶组分析和系统生物学来识别和克服ECM介导的耐药机制。
