A High Throughput Human Tumor Modeling Technology for Cancer Drug Discovery

用于癌症药物发现的高通量人体肿瘤建模技术

• 批准号: 10161750
• 负责人: Gary D Luker
• 金额: $ 36.5万
• 依托单位: UNIVERSITY OF AKRON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10161750
• 关键词: 3-Dimensional; Address; Animals; Antineoplastic Agents; Biological Models; Biopolymers; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer cell line; Carcinoma; Cells; Coculture Techniques; Color; Combined Modality Therapy; Complex; Devices; Disease model; Drops; Drug Screening; Drug resistance; Engineering; Environment; Epithelial; Event; Extracellular Matrix; Fibroblasts; Growth; Human; Immersion; In Vitro; Industrialization; Malignant Neoplasms; Mammary Neoplasms; Mediating; Microfluidic Microchips; Modeling; Molecular; Mus; Neoplasm Metastasis; Outcome; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase; Physiological; Polymers; Primary Cell Cultures; Property; Proteins; Reproducibility; Research; Resistance; Robotics; Signal Pathway; Signal Transduction; Speed; Stromal Cells; Stromal Neoplasm; System; Techniques; Technology; Testing; Tissues; Treatment Efficacy; Variant; Xenograft Model; angiogenesis; aqueous; base; bioluminescence imaging; cancer cell; cancer drug resistance; cancer therapy; chemotherapy; combinatorial; design; drug development; drug discovery; drug sensitivity; drug testing; drug use screening; effective therapy; effectiveness validation; flexibility; high throughput screening; high throughput technology; high-throughput drug screening; human model; improved; improved outcome; in vivo; inhibitor/antagonist; malignant breast neoplasm; mechanical properties; mouse model; new technology; novel; operation; patient derived xenograft model; personalized cancer therapy; physiologic model; precision drugs; precision medicine; precision oncology; response; small molecule; three dimensional cell culture; tool; triple-negative invasive breast carcinoma; tumor; tumor growth; tumor progression

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.

项目摘要 肿瘤基质，包括细胞外基质（ECM）和细胞，基本上调节 肿瘤生长和转移。癌细胞，基质细胞和ECM之间的信号传导促进 癌细胞的增殖和耐药性等其他关键结果。因此，破坏了基质 - 癌细胞信号传导对于恢复癌细胞的药物敏感性至关重要 患者。尽管有这种认可，但缺乏生理，高吞吐量人类肿瘤模型 阻碍了针对肿瘤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动物技术来重新创建这种需求 天然肿瘤的复杂性和对肿瘤信号传导的药物测试。这项便利的技术 基于用户定义的癌细胞，基质细胞和ECM的两步机器人微图解 1536 MICROWELL板中的聚合物两相系统。在 水纳米旋风在微孔的底部定居，并从浸入水相中不混溶。 然后将第二个包含基质成分的水滴分配以与纳米旋风合并 并包围癌细胞块，自发孵育时会产生微动物。这种方法 独特地提供了结合组织特异性基质蛋白和不同基质细胞的灵活性 在体内繁殖肿瘤的物理化学特性。我们将使用三重负面验证这项技术 乳腺癌（TNBC）作为疾病模型，证明了与癌相关成纤维细胞（CAFS）的影响 和ECM关于癌细胞增殖和药物反应。有了这项技术，我们将测试 破坏针对TNBC细胞治疗功效的肿瘤 - 核信号传导。这些研究将使用 TNBC细胞系和有条件重编程的细胞的工程肿瘤模型 转移性TNBC患者的癌细胞建立使用我们的TMT模型系统的可行性 精度肿瘤学。我们将使用标准化学治疗药进行组合筛查和 分子抑制剂对特定TNBC患者细胞中活性的信号通路的分子抑制剂抑制基质 - 癌细胞的介导的增殖和耐药性，并验证小鼠中最有效的治疗方法 人类TNBC的异种移植模型。通过这项研究，我们希望将我们的TMT技术建立为 将在药物发现，乳房的机理研究中广泛实施的变革性进步 癌症和其他恶性肿瘤以及精密医学。