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，基本上调节肿瘤的所有方面。 肿瘤生长和转移。肿瘤中癌细胞、基质细胞和ECM之间的信号传导促进 癌细胞增殖和耐药性等关键结果。因此，破坏基质- 癌细胞信号传导对于恢复癌细胞的药物敏感性和改善治疗结果至关重要。 患者尽管认识到了这一点，但缺乏生理性的、高通量的人类肿瘤模型， 阻碍了靶向肿瘤-基质相互作用的药物开发和发现工作。 我们将通过开发一种高通量肿瘤微组织技术来重建肿瘤细胞， 本发明的目的在于降低天然肿瘤的复杂性，并使针对肿瘤基质信号传导的药物测试成为可能。这项简单的技术 是基于两步机器人微图案化的用户定义的癌细胞，基质细胞，和ECM使用 在1536微孔板中的聚合物水性两相系统。癌细胞的3D质量在一个 水性纳米滴沉降在微孔的底部并且与浸没水相不混溶。 然后分配含有基质组分的第二水性液滴以与纳米液滴合并 并包围癌细胞团以在孵育时自发产生微组织。这种方法 独特地提供了将组织特异性基质蛋白和不同基质细胞结合到 在体内重现肿瘤的物理化学性质。我们将使用三重否定来验证这项技术 乳腺癌（TNBC）作为疾病模型，证明癌相关成纤维细胞（CAF）的作用 和ECM对癌细胞增殖和药物反应的影响。有了这项技术，我们将测试 破坏肿瘤-基质信号传导对TNBC细胞的治疗功效。这些研究将使用 TNBC细胞系和条件性重编程细胞两者的工程化肿瘤模型， 转移性TNBC患者的癌细胞，以建立使用我们的TMT模型系统用于 精准肿瘤学我们将使用标准化疗药物进行组合药物筛选， 针对在特定TNBC患者的细胞中有活性的信号传导途径的分子抑制剂，以抑制间质- 介导的癌细胞增殖和耐药性，并在小鼠中验证最有效的治疗方法 人TNBC的异种移植模型。通过这项研究，我们希望建立我们的TMT技术作为一个 将广泛应用于药物发现、乳腺癌机制研究 癌症和其他恶性肿瘤，以及精准医疗。

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 1536 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 drug responses of cancer cells. With this technology, we will test effects of disrupting tumor-stromal signaling on treatment efficacy against TNBC cells. These studies will use engineered tumor models of both TNBC cell lines and conditionally reprogrammed cells generated from cancer cells of patients with metastatic TNBC to establish the feasibility of using our TMT model system for precision oncology. We will perform combinatorial drug screening using standard chemotherapeutics and molecular inhibitors against signaling pathways active in cells of specific TNBC patients to inhibit stroma- mediated proliferation and drug resistance of cancer cells, and validate the most effective treatments in mouse xenograft models of human TNBC. Through this research, we expect to establish our TMT technology as a transformative advance that will be implemented broadly for drug discovery, mechanistic studies of breast cancer and other malignancies, and precision medicine.