Harnessing human brain and liver microphysiological systems for testing therapeutics for metastatic melanoma

项目摘要 评估药物治疗转移性黑色素瘤疗效的标准是患者的反应， 但临床上迫切需要一种能够支持药物疗效预测的人类替代模型， 从而将患者从反复尝试的治疗中拯救出来，这最终将成为指导 选择针对患者的药物疗法。今天，患者派生的使用受到了极大的关注 异种移植(PDX)，将患者的肿瘤移植到免疫缺陷小鼠体内，以在 用老鼠制作病人肿瘤的模型。不幸的是，这个过程缓慢且昂贵，并且基于 动物的微环境而不是人类的微环境。微生理系统(MPS)，包括 芯片上的器官、组织芯片和工程有机体，可以使用人类细胞来构建 体外微环境。拟议的研究将建立在范德比尔特大学强大的合作基础上 大学、匹兹堡大学和威斯康星大学共同开发强大的议员来解决 需要建立病人对癌症治疗反应的模型。这个项目将研究组织如何 微环境影响转移性黑色素瘤细胞的生长及其对药物的反应 范德比尔特神经血管单位组织芯片、匹兹堡芯片上肝脏和威斯康星工程有机体 对于大脑和肝脏，每一种都包括多种细胞类型。这项研究的重点将放在 转移性级联--肿瘤细胞在远离原发灶的部位生长。这种增长是由 通过“种子与土壤”的相互作用，肿瘤的“种子”与组织微环境的“土壤”。而不是 利用老鼠作为土壤，患者的癌细胞将被种植到大脑和肝脏MPS提供的土壤中 从人类诱导的多能干细胞衍生的结构。目标是1)实施一套共同的 人类器官结构(芯片上的肝脏、神经血管单元和来自单个人类干细胞的工程有机体 细胞来源)，2)展示了这些人体器官结构的成功种植以及转移的皮肤 来自范德比尔特和匹兹堡患者的黑色素瘤或葡萄膜黑色素瘤细胞，以及3)比较 植入芯片上器官并经工程处理的患者癌细胞对药物的反应 通过现有的PDX系列对相同药物产生反应的有机化合物。该项目将为以下方面提供指导 哪种体外人体模型在移植到临床时可能更能预测患者的结果，基于 部分取决于肿瘤的类型、患者样本的性质和患者的基因。它还将测试 假设范德比尔特、匹兹堡和威斯康星州开发的人类MPS设备和模型将 为研究肿瘤转移提供了更真实的、体外的三维人体微环境 鼠标PDX。最后阶段将是精确医学的概念验证演示，其中 大脑和肝脏的微环境可能来自患者诱导的多能干细胞。

Project Summary The standard for assessing the effectiveness of drugs to treat metastatic melanoma is the patient's response, but there is a pressing clinical need for a human surrogate model that could support prediction of drug efficacy, thereby saving the patient from trial and error treatments, and that would ultimately serve as a guide for the selection of patient-targeted drug therapies. Today, there is significant interest in the use of patient-derived xenografts (PDXs), in which a patient's tumor is implanted into an immune-deficient mouse, to create in the mouse a model of the patient's tumor. Unfortunately, this process is slow and expensive and is based upon an animal microenvironment rather than a human one. Microphysiological systems (MPS), which encompass organs-on-chips, tissue chips, and engineered organoids, can be constructed using human cells to create an in vitro microenvironment. The proposed research would build upon a strong collaboration at Vanderbilt University, the University of Pittsburgh, and the University of Wisconsin to develop powerful MPS to address the need for models of a patient's response to cancer therapy. This project will study how the tissue microenvironment affects the growth of metastatic melanoma cells and their response to drugs by using the Vanderbilt neurovascular unit tissue chip, the Pittsburgh liver-on-chip, and the Wisconsin engineered organoids for brain and liver, each of which includes multiple cell types. The research will focus on the final stage in the metastatic cascade – the growth of tumor cells at sites distant from the primary tumor. This growth is governed by “seed and soil” interaction between the tumor “seed” and the tissue microenvironment “soil.” Instead of using a mouse as the soil, patients' cancer cells will be planted into the soil provided by brain and liver MPS constructs derived from human induced pluripotent stem cells. The aims are 1) Implement a common set of human organ constructs (liver-on-chip, neurovascular unit, and engineered organoid from a single human stem cell source), 2) Demonstrate successful seeding of these human organ constructs with metastatic cutaneous melanoma or uveal melanoma cells derived from Vanderbilt and Pittsburgh patients, and 3) Compare the response to drugs by patients' cancer cells that have been seeded into the organs-on-chips and engineered organoids with the response to the same drugs by existing PDX lines. This project will provide guidance as to which in vitro human model might be more predictive of patient outcome when translated to the clinic, based in part upon the type of tumor, the nature of the patient sample, and the patient genotype. It will also test the hypothesis that the human MPS devices and models developed at Vanderbilt, Pittsburgh, and Wisconsin will provide a more realistic, in vitro, three-dimensional human microenvironment to study tumor metastasis than mouse PDXs. The final phase will be a proof-of-concept demonstration of precision medicine in which the microenvironment of the brain and liver could be from the patient's induced pluripotent stem cells.