Organoid modeling of pre-metastatic niche formation in the liver by primary colorectal tumor secreted factors

原发性结直肠肿瘤分泌因子在肝脏中形成转移前生态位的类器官模型

基本信息

批准号：
10355869
负责人：
Aleksander Skardal
金额：
$ 21.81万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-23 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10355869
关键词：
3-Dimensional Affect Animal Model Antineoplastic Agents Architecture Automobile Driving Biocompatible Materials Biological Biological Markers Biomedical Engineering Blood Circulation Bone Marrow Cancer Etiology Cause of Death Cell Membrane Permeability Cell Proliferation Cells Cessation of life Clinical Collagen Collagen Fiber Colorectal Cancer Colorectal Neoplasms Development Devices Diagnosis Disease Distant Drug Targeting Engraftment Environment Event Extracellular Matrix Extravasation Fibronectins Future Goals Growth Hematopoietic stem cells Hepatic Hepatocyte Homing Human Hydrogels Immune Immune system In Vitro Inflammatory Interruption Intervention Kinetics Kupffer Cells Laminin Lead Link Liquid substance Liver Measures Mediator of activation protein Metastatic Neoplasm to the Liver Metastatic to Methodology Microfluidic Microchips Microfluidics Modeling Motivation Neoplasm Metastasis Organ Organoids Paper Patients Phenotype Population Primary Neoplasm Process Publishing Resources Signal Transduction Site Stromal Cells Targeted Research Technology Testing Therapeutic Intervention Tissue Engineering Tissue Model Tissues Tumor Cell Line Tumor Tissue Tumor-Derived base cancer invasiveness cancer therapy chemotherapy clinically relevant colorectal cancer metastasis cytokine effective therapy in vitro Model in vivo Model inflammatory marker innovation liver inflammation mechanical properties mesenchymal stromal cell metastatic process migration mimetics mortality neoplastic cell new therapeutic target novel novel strategies operation paracrine pre-clinical prevent recruit response risk prediction stellate cell time interval tool transcriptome sequencing tumor tumor microenvironment

项目摘要

Metastatic disease remains the primary cause of cancer-related deaths. In colorectal cancer (CRC), the liver is the primary site of metastases; and, the majority of patients diagnosed with CRC die as a result of their hepatic metastases. This provides a powerful motivation to study the mechanisms behind the complicated metastatic cascade, hopefully leading to novel approaches for intervention. However, studying the biological mechanisms that contribute to metastatic disease has been difficult. One such target for research that has arisen recently is the pre-metastatic niche (PMN), which forms distant from the primary tumor, but creates favorable environments for metastasizing cells to take hold. Studying the PMN is challenging, as in animal models it is nearly impossible to predict when changes might happen in the PMN, and after identification of metastases, it is too late. A clinically-relevant in vitro model in which metastasis and the changes to tissues leading up to metastasis can be observed and manipulated would be an invaluable resource for probing these mechanisms and for identifying ways to circumvent them. Our team has now published several papers describing our bioengineered in vitro metastasis-on-a-chip (MOC) platform that is comprised of microfluidic devices and 3D extracellular matrix (ECM) hydrogel-based tumor and tissue organoids. Our objective is to use these tools to identity and pinpoint early changes to the liver PMN that occur because of primary tumor-secreted mediators and assess their contributions to CRC metastasis. We will accomplish this by employing 3D liver, CRC, and bone marrow hematopoietic progenitor cell (BM-HPC) organoids, contained in a microfluidic platform to quantify liver remodeling and BM- HPC recruitment, and in turn, how this remodeling influences the likelihood and kinetics of in vitro metastasis. We hypothesize that within our novel bioengineered in vitro platform, secreted mediators from tumor cell line organoids and patient-derived tumor organoids (PTOs) of varying grade will induce a corresponding spectrum of ECM remodeling, architectural dysregulation, and BM-HPC recruitment, which when heightened, corresponds with increased metastasis. In Aim 1, we will subject primary human liver and BM-HPC organoids to CRC conditioned media, measure hepatic stromal (stellate) and immune (Kupffer) cell activation, and quantify ECM remodeling in terms of composition, architecture, and mechanical properties. As part of this aim, we will dissect out the relative contributions of stellate cells, Kupffer cells, and BM-HPCs to these changes. In Aim 2, our MOC device will include tumor organoids in addition both the liver and BM-HPC organoids and we will assess PMN formation in this dynamic multi-tissue platform. Metastasis from the tumor to liver organoid will be measured. This will enable us to link changes of the PMN to metastatic events. Ultimately, these studies will provide opportunities to identify tumor cell secreted mediators and distinct features of the PMN that could potentially serve as future targets of the early metastatic process prior to becoming incurable as well as biomarkers for prediction of metastasis in patients.

转移性疾病仍然是癌症相关死亡的主要原因。在结直肠癌 (CRC) 中，肝脏是转移的原发部位；而且，大多数被诊断患有结直肠癌的患者死于肝脏疾病转移。这为研究复杂转移背后的机制提供了强大的动力。级联，有望带来新的干预方法。然而，研究其生物学机制导致转移性疾病的方法一直很困难。最近出现的这样一个研究目标是转移前生态位（PMN），远离原发肿瘤，但创造了有利的环境让转移细胞扎根。研究 PMN 具有挑战性，因为在动物模型中这几乎是不可能的预测 PMN 何时可能发生变化，而在识别出转移后，为时已晚。一个临床相关的体外模型，其中可以观察转移和导致转移的组织变化观察和操纵将是探索这些机制和识别这些机制的宝贵资源。规避它们的方法。我们的团队现已发表多篇论文，描述我们的体外生物工程由微流体装置和 3D 细胞外基质 (ECM) 组成的片上转移 (MOC) 平台基于水凝胶的肿瘤和组织类器官。我们的目标是使用这些工具尽早识别和查明由于原发性肿瘤分泌介质而发生的肝脏 PMN 变化并评估其贡献至结直肠癌转移。我们将通过采用 3D 肝脏、CRC 和骨髓造血来实现这一目标祖细胞 (BM-HPC) 类器官，包含在微流体平台中，用于量化肝脏重塑和 BM- HPC 招募，以及这种重塑如何影响体外转移的可能性和动力学。我们假设在我们的新型生物工程体外平台中，肿瘤细胞系分泌的介质不同等级的类器官和患者来源的肿瘤类器官（PTO）将产生相应的谱 ECM 重塑、结构失调和 BM-HPC 募集，当其增强时，对应随着转移的增加。在目标 1 中，我们将对原代人类肝脏和 BM-HPC 类器官进行 CRC 条件培养基，测量肝基质（星状）和免疫（Kupffer）细胞活化，并量化 ECM 在成分、结构和机械性能方面进行重塑。作为这一目标的一部分，我们将剖析找出星状细胞、库普弗细胞和 BM-HPC 对这些变化的相对贡献。在目标 2 中，我们的 MOC 设备将包括肿瘤类器官以及肝脏和 BM-HPC 类器官，我们将评估 PMN 在这个动态的多组织平台上形成。将测量从肿瘤到肝类器官的转移。这将使我们能够将 PMN 的变化与转移事件联系起来。最终，这些研究将提供识别肿瘤细胞分泌介质和 PMN 独特特征的机会作为无法治愈之前早期转移过程的未来目标以及生物标志物预测患者的转移。