Engineered microtumor arrays for development of combination therapies

项目摘要 恶性黑色素瘤是产生色素的黑色素细胞的肿瘤，并且是导致大多数恶性黑色素瘤的原因。 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，存活率就会大大降低。最近的一些研究 已经表明，在某些情况下，来自肿瘤微环境的线索可以表观遗传重编程， 将黑素瘤细胞转化为恶性黑素瘤起始细胞（MIC），所述恶性黑素瘤起始细胞是耐药性的并且为侵袭做好准备， 转移在Kilian实验室中，我们发现了肿瘤拓扑结构与肿瘤细胞活化之间的关系。 一种致瘤性MIC，可能证明是转移前的早期转化步骤。在科普兰实验室， 我们已经开发了新的联合疗法来治疗黑色素瘤和几种患者来源的肿瘤异种移植物， （PDTX）模型，其准确地模拟患者对标准护理的反应。我们将雇用我们的一套 工程细胞外基质，以破译拓扑结构，力学和基质之间的相互作用， 本发明还提供了组合物在引导显示不同程度的药物抑制的患者来源的细胞中的MIC状态的活化中的用途。 灵敏度我们将这些矩阵参数转化为一种新的三维几何结构的组织工程 微肿瘤模型为了评估其作为药物开发工具的潜力，我们将制作一个96孔板- 基于格式，并评估微肿瘤对标准治疗和前瞻性新疗法的反应。我们将使用 原位异种移植在无胸腺裸鼠研究我们的细胞的生长，侵袭和传播，并使用这个 这些信息为我们的3D肿瘤模拟模型的设计提供了信息。利用我们的微肿瘤阵列 治疗开发，我们将使用我们的肿瘤模拟物进行一项联合治疗的小型试点研究， 在体外和作为一种新的体内微肿瘤异种移植物-辨别我们的仿生系统如何密切重演 与细胞系和异种移植物相比，肿瘤发生和药物敏感性。该项目旨在建立一个 补充或甚至替代患者来源的异种移植物（PDX）模型的方法，其中患者的细胞 可将来自活组织检查或切除的肿瘤细胞整合到肿瘤模拟物中用于个体化药物。

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.