Development of 3D organ-specific models of colorectal cancer metastasis

描述（申请人提供）：了解癌症转移的生物学对改善癌症的治疗至关重要。在这些努力中的一个关键挑战是缺乏易于使用的肿瘤模型，可以概括转移性疾病的状况或过程。目前的模型要么难以研究，要么无法复制肿瘤转移的复杂微环境。我们的申请旨在通过应用组织工程的最新进展来解决对癌症转移模型的需求。组织工程最近的一个突破是脱细胞组织的发展。里德博士开发的一种用于生成脱细胞组织的新技术，除了保留细胞外基质结合的生长因子和细胞因子。使用这种技术产生的脱细胞组织被称为生物基质支架。里德小组已经证明，生物基质支架在化学和功能上都是组织特异性的，而不是物种特异性的。使用生物基质支架，我们已经获得了令人兴奋的初步数据。我们发现大肠癌细胞HT29、SW480和CaCO2可以在包被肝和肺生物基质支架的组织培养皿上自发形成3D菌落。更重要的是，我们已经证明了在肝和肺生物基质支架上生长的细胞对化疗和放疗的治疗反应是不同的。这种器官特异性反应尚未在其他3D培养系统中观察到。最后，我们已经证明，来自结直肠癌肝转移的人原发肿瘤细胞在体外培养的肝生物基质上形成的菌落明显多于在肺生物基质、胶原蛋白或塑料上生长的菌落。基于我们的初步数据，我们假设我们可以使用生物基质支架来生成三维的体外和离体肿瘤转移模型。在本次申请中，我们计划以大肠癌为模型疾病，建立肝、肺转移的大肠癌模型。我们推测，我们提出的模型可以概括结直肠癌转移到肝脏和肺部的生物学过程，并预测转移的治疗反应。我们的应用程序有两个特定的目标。第一个目标将专注于仅使用组织特异性生物基质支架开发结直肠癌转移的体外器官特异性3D模型。我们的第二个目标是利用肝生物基质支架的再细胞化制备的肝类器官，开发结直肠癌肝转移的3D离体模型。我们研究的成功将导致新的体外/离体癌症转移模型的发展，从而更好地模拟疾病过程。这些可以成为研究转移生物学的有力工具，包括：转移机制；物理力在转移中的作用；以及控制转移潜能的基质成分的鉴定。此外，模型可用于针对癌症转移到特定器官的体外治疗筛选试验。我们的策略也可以应用于其他类型的癌症和转移到其他器官。

DESCRIPTION (provided by applicant): Understanding the biology of cancer metastases is critical to improving the treatment of cancer. A key challenge in these efforts has been the lack of easy-to-use tumor models that can recapitulate the metastatic disease condition or process. Current models are either too difficult to study or unable to replicate the complex microenvironment of tumor metastasis. Our application aims to address the need for models of cancer metastasis by applying recent advances in tissue engineering. A recent breakthrough in tissue engineering has been the development of decellularized tissue. One novel technique for generating decellularized tissue, developed by Dr. Reid, preserves growth factors and cytokines that are matrix-bound in addition to the extracellular matrix. Decellularized tissue generated using this technique has been termed biomatrix scaffolds. The Reid group has shown that biomatrix scaffolds are tissue-specific but not species-specific both chemically and functionally. Using biomatrix scaffolds, we have obtained exciting preliminary data. We have found that colorectal cancer cells, HT29, SW480 and CaCO2, can spontaneously form 3D colonies on tissue culture dishes coated with liver and lung biomatrix scaffolds. More importantly, we have demonstrated that treatment responses to chemotherapy and radiotherapy are different between cells grown on liver and lung biomatrix scaffolds. Such organ-specific responses have not been observed with other 3D culture systems. Lastly, we have shown that human primary tumor cells from hepatic metastases of colorectal cancer form significantly more colonies when grown on liver biomatrix in vitro compared to that on lung biomatrix, collagen or plastic. Based on our preliminary data, we hypothesize that we can use biomatrix scaffolds to generate 3D in vitro and ex vivo models of cancer metastasis. In this application, we plan to use colorectal cancer as a model disease and develop models of colorectal cancer with liver and lung metastases. We theorize that our proposed models can recapitulate the biology of colorectal cancer metastasis to liver and lung as well as predict treatment responses of metastases. Our application has two specific aims. The first aim will focus on the development of in vitro organ-specific 3D models of colorectal cancer metastasis using tissue-specific biomatrix scaffolds only. Our second aim will focus on the development of 3D ex vivo models of colorectal cancer liver metastases using liver organoids prepared by recellularization of liver biomatrix scaffolds. Success with our research can lead to the development of novel in vitro/ex vivo models of cancer metastasis that can better mimic the disease process. These can become powerful tools for studying the biology of metastasis including: mechanisms of metastasis; roles of physical forces on metastasis; and identification of matrix components controlling metastatic potential. Furthermore, models can be useful for in vitro therapeutic screening assays targeted towards cancer metastasis to a specific organ. Our strategy can also be applied to other types of cancers and metastasis to other organs.