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

项目摘要 评估治疗转移性黑色素瘤的药物有效性的标准是患者的反应， 但是临床上迫切需要一种能够支持药物功效预测的人类替代模型， 从而使患者免于试错治疗，这最终将成为 选择针对患者的药物治疗。今天，对使用患者来源的 异种移植物（PDX），其中将患者的肿瘤植入免疫缺陷小鼠中，以在小鼠体内产生肿瘤。 老鼠是病人肿瘤的模型。不幸的是，这个过程是缓慢和昂贵的，是基于一个 动物微环境而不是人类微环境。微生理系统（MPS），包括 芯片上的器官、组织芯片和工程化的类器官可以使用人类细胞来构建， 体外微环境拟议中的研究将建立在范德比尔特强有力的合作基础上 美国匹兹堡大学和威斯康星州大学合作开发强大的MPS， 对病人对癌症治疗反应的模型的需求。这个项目将研究组织如何 微环境影响转移性黑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，匹兹堡肝脏芯片和威斯康星州工程类器官 对于大脑和肝脏，每一个都包括多种细胞类型。研究将集中在最后阶段， 转移级联-肿瘤细胞在远离原发肿瘤的部位生长。这种增长是由 通过肿瘤"种子"和组织微环境"土壤"之间的"种子和土壤"相互作用。而不是 以老鼠为土壤，病人的癌细胞将被植入大脑和肝脏MPS提供的土壤中。 来源于人诱导多能干细胞的构建体。目标是：（1）实施一套共同的 人类器官构建体（肝芯片、神经血管单元和来自单个人类干的工程化类器官 2）证明这些人器官构建体与转移性皮肤癌细胞的成功接种， 黑色素瘤或葡萄膜黑色素瘤细胞来源于范德比尔特和匹兹堡患者，和3）比较 病人的癌细胞对药物的反应，这些癌细胞已经被接种到芯片上的器官中， 类器官对现有PDX细胞系的相同药物有反应。该项目将提供指导， 当转化到临床时，哪种体外人体模型可能更能预测患者的结局， 部分取决于肿瘤的类型、患者样品的性质和患者基因型。它还将测试 假设在范德比尔特、匹兹堡和威斯康星州开发的人类MPS设备和模型将 提供了一个更现实的，在体外，三维人体微环境，研究肿瘤转移比 小鼠PDX。最后阶段将是精准医疗的概念验证演示， 大脑和肝脏的微环境可能来自患者的诱导多能干细胞。

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.