A High Throughput Human Tumor Modeling Technology for Cancer Drug Discovery

项目摘要 肿瘤间质，包括细胞外基质(ECM)和细胞，基本上调节所有方面的 肿瘤的生长和转移。肿瘤中癌细胞、间质细胞和细胞外基质之间的信号转导促进 除其他关键结果外，还包括癌细胞的增殖和耐药性。因此，破坏基质- 癌细胞信号转导对恢复癌细胞的药物敏感性和改善预后至关重要 病人。尽管认识到这一点，但缺乏生理学的、高通量的人类肿瘤模型 阻碍针对肿瘤-间质相互作用的药物开发和发现努力。 我们将通过开发高通量肿瘤微组织技术来满足这一需求，以重建 此外，我们还研究了天然肿瘤的复杂性，并实现了针对肿瘤间质信号的药物测试。这种轻便的技术 基于用户定义的癌细胞、间质细胞和ECM的两步机器人微图案化，使用 微孔板中的聚合物双水相体系。在水溶液中形成3D癌细胞团块 纳米液滴沉淀在微孔底部，与浸泡水相不相容。一秒钟 然后分配含有基质成分的水滴以与纳米液滴合并并包围 该癌细胞团在孵育时自发地产生微组织。这种方法独特地提供了 将组织特异性基质蛋白和不同的基质细胞结合起来进行复制的灵活性 体内肿瘤的物理化学性质。我们将使用三重负乳房来验证这项技术 癌症(TNBC)作为疾病模型，展示了癌症相关成纤维细胞(CAF)和 ECM对癌细胞增殖和基质侵袭的影响。这些研究将使用工程化的肿瘤模型 TNBC细胞系和患者来源的CAF来建立我们的肿瘤模型的可行性 特定于患者的细胞。通过这项研究，我们希望建立我们的肿瘤模型作为一种变革性的 这一进展将广泛应用于乳腺癌的药物发现和机制研究。

Project Summary Tumor stroma, encompassing both extracellular matrix (ECM) and cells, regulates essentially all aspects of tumor growth and metastasis. Signaling among cancer cells, stromal cells, and ECM in tumors promotes proliferation of cancer cells and drug resistance among other key outcomes. Therefore, disrupting stroma- cancer cells signaling is essential to restoring drug sensitivity of cancer cells and improving outcomes for patients. Despite this recognition, the lack of physiologic, high throughput human tumors models significantly impedes drug development and discovery efforts targeting tumor-stromal interactions. We will address this need by developing a high throughput tumor microtissue technology to recreate the complexity of native tumors and enable drug testing against tumor-stromal signaling. This facile technology is based on two-step robotic micropatterning of user-defined cancer cells, stromal cells, and ECM using a polymeric aqueous two-phase system in microwell plates. A 3D mass of cancer cells is formed in an aqueous nanodrop settled at the bottom of a microwell and immiscible from the immersion aqueous phase. A second aqueous drop containing the stromal components is then dispensed to merge with the nanodrop and surround the cancer cell mass to spontaneously generate a microtissue upon incubation. This approach uniquely offers the flexibility of incorporating tissue-specific matrix proteins and different stromal cells to reproduce physicochemical properties of tumors in vivo. We will validate this technology using triple negative breast cancer (TNBC) as a disease model, demonstrating effects of carcinoma-associated fibroblasts (CAFs) and ECM on proliferation and matrix invasion of cancer cells. These studies will use engineered tumor models of both TNBC cell lines and patient-derived CAFs to establish the feasibility of tailoring our tumor model to patient-specific cells. Through this research, we expect to establish our tumor model as a transformative advance that will be implemented broadly for drug discovery and mechanistic studies of breast cancer.