An organotypic model recapitulating colon cancer microenvironment and metastasis

项目总结 此U01是对FOA PAR-16-105癌症组织工程协作性：启用的响应 用于癌症研究的仿生组织工程技术。在这篇文章中，我们将解决 我们对转移性结直肠癌(CRC)的发生和发展的了解。这一差距在很大程度上是应该的。 部分原因是，目前的体外、体外甚至体内研究模型很难建立或拥有 适用性有限。因此，我们的目标是展示三种创新的转移模型的实用性。 (长期器官表型模型和使用多器官微流控装置的模型)和新的体内模型 (免疫活性囊胚模型)。利用这些模型，我们将研究交互式细胞分泌体， 原发和转移成纤维细胞肿瘤微环境的分化、迁移和侵袭 以确定结直肠癌转移的关键因素。我们的假设是：炎症相关的成纤维细胞和 代谢条件的改变通过转录调节促进上皮转移。根据我们最近的调查 新模型的成功设计，我们提出了三个目标。目的1.将分泌细胞因子的基质整合到 我们的器官模型，以研究炎症对肿瘤干细胞(SCs)和 侵袭/迁移细胞表型。目的2.设计一种使用微流体体的肝转移模型。 研究转移细胞转录组和表观遗传重编程的芯片平台。目标3.验证我们的入网 体外模型采用体内免疫熟练的小鼠转移模型。在目标1中，脱细胞的人类结肠 将从基质中重新植入正常或癌变的成纤维细胞，并以 了解这些操作如何影响该模型中的侵袭和分化。 为了补充细胞的发现，我们将使用RNA-seq和atac-seq来识别转录组的变化。 序列号。Aim 2使用创新的Body-on-a芯片微流控设备测试自分泌和旁分泌增殖性 以及在有利于肿瘤进展的条件下的分泌反应，包括低氧和低糖。 与目标1一样，不同环境中的上皮和成纤维细胞将被转录。Aim 3使用了一种新的 具有免疫活性的囊胚模型，并将经历相同的转录图谱。所有3款车型都将是 使用比较、信息学的方法进行链接。这种方法是创新的，因为它使用了人类的结肠组织 所有目标和问题的转录贡献和景观的可及染色质在每个模型。 我们的多学科团队包括一名结直肠外科医生兼科学家和具有以下专业知识的生物医学工程师 癌症干细胞，信号，转移，器官类型和芯片上器官平台。调查结果将是 意义重大，因为他们将建立改进的方法来研究晚期CRC的发病机制，并 开发患者衍生的高通量细胞模型以测试治疗干预措施。

PROJECT SUMMARY This U01 is in response to FOA PAR-16-105 Cancer Tissue Engineering Collaborative: Enabling Biomimetic Tissue-Engineered Technologies for Cancer Research. In it we will address the substantial gap in our knowledge of the initiation and progression of metastatic colorectal cancer (CRC). This gap is due, in large part, to the fact that current in vitro, ex vivo, and even in vivo research models are hard to establish or have limited applicability. Accordingly, our objectives are to show the utility of three innovative models of metastasis (a long-term organotypic model and a model that uses multi-organ microfluidic devices) and a new in vivo model (immunocompetent blastocyst model). Using these models, we will investigate the interactive cellular secretome, differentiation, migration and invasion of the primary and metastatic fibroblast tumor microenvironment in order to identify critical contributions to CRC metastasis. Our hypothesis is: inflammation-associated fibroblasts and altered metabolic conditions promote epithelial metastasis via transcriptional regulation. Based on our recent successful engineering of novel models, we propose 3 Aims. Aim 1. To integrate cytokine-secreting stroma into our organotypic model in order to study the influence of inflammation on cancer stem cells (SCs) and on invasion/migration cell phenotypes. Aim 2. To engineer a liver metastasis model using a microfluidic body-on-a- chip platform to study transcriptome and epigenetic reprogramming of metastatic cells. Aim 3. To validate our in vitro models using in vivo immunoproficient murine metastatic models. In Aim 1, decellularized human colons will be repopulated with normal or cancerous fibroblasts from the stroma, and cancer epithelial cells in the form of cancer SCs and organoids to ask how these manipulations influence invasion and differentiation in this model. To complement the cellular findings, we will identify alterations in the transcriptome using RNA-seq and ATAC- seq. Aim 2 uses the innovative body-on-a chip microfluidic device to test autocrine and paracrine proliferative and secretory responses under conditions that favor oncogenic progression, including hypoxia and low glucose. As in Aim 1, epithelia and fibroblasts in the different milieus will be transcriptionally profiled. Aim 3 uses a new immunocompetent blastocyst model and will undergo the same transcriptional profiling. All 3 models will be linked using comparative, informatics approaches. The approach is innovative as it uses human colon tissues in all Aims and asks about the transcriptional contribution and landscape of accessible chromatin in each model. Our multidisciplinary team includes a colorectal surgeon-scientist and biomedical engineers with expertise in cancer SCs, signaling, metastases, and organotypic and organ-on-a-chip platforms. The findings will be significant, as they will establish improved approaches for studying the pathogenesis of advanced CRC, and for developing patient-derived, high-throughput cell models to test therapeutic interventions.