Engineered microtumor arrays for development of combination therapies

项目总结 恶性黑色素瘤是一种产生色素的黑素细胞的肿瘤，它是大多数 与皮肤癌相关的死亡。皮肤黑色素瘤可以通过手术切除成功治疗； 然而，一旦疾病转移，存活率就会显著降低。最近的一些研究 已经提出，在某些情况下，来自肿瘤微环境的线索可以在表观遗传上重新编程 黑色素瘤细胞转化为恶性黑色素瘤启动细胞(MIC)，这种细胞具有抗药性，并为侵袭和 转移。在基利安实验室，我们发现了肿瘤拓扑和激活的关系 一种致瘤的MIC，可能是转移前的早期转化步骤。在科普兰实验室， 我们已经开发了治疗黑色素瘤的新的联合疗法，以及几个患者衍生的肿瘤异种移植。 (PDTX)模型，准确模拟患者对标准护理的反应。我们将雇佣我们的套间 设计细胞外基质以破译拓扑、力学和基质之间的相互作用 在显示不同程度药物的患者来源细胞中引导MIC状态激活的组合物 敏感度。我们将把这些矩阵参数转换成一种新的3D几何结构的组织工程 微瘤模型。为了评估作为药物开发工具的潜力，我们将制造一个96孔板- 基于形成和评估微肿瘤对标准护理和预期新疗法的反应。我们将使用 在裸鼠体内进行原位异种移植来研究我们的细胞的生长、侵袭和扩散，并使用这个 为我们的3D肿瘤模拟模型的设计提供信息。利用我们的微肿瘤阵列 在治疗开发方面，我们将使用我们的肿瘤模拟药物对联合疗法进行一项小型试验性研究- 在体外和作为一种新的微型肿瘤异种移植-识别我们的仿生系统有多接近重演 与细胞系和异种移植相比的致瘤性和药物敏感性。该项目旨在建立一个 患者来源的异种移植(PDX)模型的补充甚至替代方法，其中患者的细胞 从活组织检查或切除中衍生出来的可能被集成到个体化药物的肿瘤模拟中。

PROJECT SUMMARY Malignant melanoma is a tumor of the pigment-producing melanocytes, and is responsible for the majority of skin cancer related deaths. Cutaneous melanoma can be successfully treated through surgical excision; however, once the disease has metastasized, the survival rate is significantly reduced. Some recent studies have suggested that in certain contexts, cues from the tumor microenvironment can epigenetically reprogram melanoma cells into a malignant melanoma initiating cell (MIC) that is drug resistant and primed for invasion and metastasis. In the Kilian laboratory we have discovered a relationship between tumor topology and activation of a tumorigenic MIC that may prove an early transformation step preceding metastasis. In the Copland laboratory, we have developed novel combination therapies to treat melanoma, and several patient derived tumor xenograft (PDTX) models that accurately mimic patient response to standard of care. We will employ our suite of engineered extracellular matrices to decipher the interplay between topology, mechanics and matrix composition, in guiding activation of the MIC state in patient derived cells that display varying degrees of drug sensitivity. We will translate these matrix parameters into a novel 3D geometrically structured tissue engineered microtumor model. To evaluate the potential as a tool for drug development we will fabricate a 96-well plate- based format and evaluate microtumor response to standard of care and prospective new therapies. We will use orthotopic xenografts in athymic nude mice to study growth, invasion and dissemination of our cells, and use this information to inform the design of our model 3D tumor-mimics. Towards leveraging our microtumor array for therapeutic development, we will perform a small pilot study of combination therapies using our tumor-mimics— in vitro and as a novel in vivo microtumor xenograft—to discern how closely our biomimetic system recapitulates oncogenesis and drug sensitivity compared to cell lines and xenografts. This project aims to establish a complementary or even alternative approach to patient derived xenograft (PDX) models, where a patient’s cells derived from biopsy or excision may be integrated into a tumor-mimic for individualized medicine.