A microengineered colon cancer-chip designed to investigate tumor-stromal interactions driving cancer progression

项目总结 在美国，结直肠癌(CRC)是癌症相关死亡的第二大原因。尽管 随着新疗法的引入，转移性疾病的五年存活率保持在10%左右。至 提高我们对肿瘤进展和药物疗效的认识，建立临床前肿瘤模型至关重要 准确地反映了结直肠癌“体外”的天然病理生理学。肿瘤的复杂性 微环境(TME)，包括基质细胞类型和机械力，在现有的 临床前模型。基于微流控技术的芯片上器官技术，旨在模拟3-D 人体器官环境在机械上(例如，流体流动和循环变形)和生物化学上(例如， 营养消化、分泌、运输)，最近为研究人员提供了对 扩大了对TME的控制。在这项提案中，我们正在采用芯片上的器官技术来创造人类 结肠器官芯片植入患者来源的CRC细胞和间质元素(内皮细胞和癌症- 相关组织中的相关成纤维细胞(CAF)：具有集成微流体的组织隔室和 伸展以模拟体内蠕动样运动。我们的目标是创造一种生理上相关的器官- 依赖肿瘤模型，将允许长期培养和表征结直肠癌细胞动力学，以及 作为测试特定治疗方式以防止或延缓肿瘤进展的平台。 使用新的成像分析方法和基于组学的方法，我们将评估物理(即， 蠕动)和细胞(CAF)微环境在结肠肿瘤进展中的作用。在目标1中，我们将培养患者- 衍生的CRC-on-Chip结合原代正常和肿瘤上皮评估CRC肿瘤生长和 在周期性拉伸存在的情况下，早期转移扩散(即侵入血管通道)，模仿 蠕动。在目标2中，我们将分析间质CAF微环境对肿瘤细胞行为和 检查患者间CAF功能异质性的程度和作用。在目标3中，我们将评估药物 通过监测药物诱导的细胞反应来筛选CRC-on-Chip平台的能力，以及 确定芯片上的药物反应是否可以预测患者的临床反应。更长远的愿景 开发一种更接近人类疾病的微工程CRC模型是为了加快 识别和筛选新的药物靶点和创新的治疗策略，重点是干扰 肿瘤-间质相互作用。我们的多学科团队具有互补的专业知识，具有开创性 技术和临床材料的可用性使我们处于有利地位，可以成功地解决重要的 结直肠癌发生的方方面面，并促进我们对肿瘤生态位的理解。

PROJECT SUMMARY Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Despite the introduction of novel therapies, the five-year survival rate for metastatic disease remains around 10%. To improve our understanding of tumor progression and drug efficacy, it is vital to develop preclinical tumor models that accurately reflect the native pathophysiology of CRC “ex vivo”. The complexities of the tumor microenvironment (TME), including stromal cell types and mechanical forces, are not fully reproduced in existing preclinical models. Microfluidic-based “organ-on-chip” technologies, which are designed to simulate the 3-D human organ environment both mechanically (e.g., fluid flow and cyclic deformation) and biochemically (e.g., nutrient digestion, secretion, transport), have recently provided researchers with greater insights into and expanded control over the TME. In this proposal, we are adapting the organ-on-chip technology to create human colon organ chips colonized with patient-derived CRC cells and stromal elements (endothelial and cancer- associated fibroblast (CAF) cells) in relevant tissue:tissue compartments with integrated microfluidics and stretching to mimic in vivo peristalsis-like motions. The goal is to create a physiologically relevant, organ- dependent tumor model that will allow for long-term culture and characterization of CRC cellular dynamics, and serve as a platform for testing specific therapeutic modalities to prevent or delay tumor progression. Using novel imaging assays and “omics” based approaches, we will evaluate the role of the physical (i.e., peristalsis) and cellular (CAFs) microenvironment in colon tumor progression. In Aim 1, we will develop patient- derived CRC-on-Chips incorporating primary normal and tumor epithelium to assess CRC tumor growth and early metastatic spread (i.e., invasion into the vascular channel) in the presence of cyclic stretch, mimicking peristalsis. In Aim 2, we will analyze the impact of the stromal CAF microenvironment on tumor cell behavior and examine the extent and role of inter-patient CAF functional heterogeneity. In Aim 3, we will assess the drug screening capabilities of the CRC-on-Chip platform via monitoring of drug-induced cellular responses, and determine whether drug responses on chips can predict patient clinical responses. The longer-term vision for developing a microengineered CRC model that more closely resembles human disease is to expedite the identification and screening of novel drug targets and innovative treatment strategies with a focus on disrupting tumor-stromal interactions. Our multi-disciplinary team with complementary expertise, the groundbreaking technologies, and availability of clinical materials put us in a prime position to successfully address important aspects of CRC tumorigenesis and advance our understanding of the tumor niche.