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

• 批准号: 10686798
• 负责人: Aleksander Skardal
• 金额: $ 17.75万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 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/10686798
• 关键词: 3-Dimensional; Affect; Animal Model; Antineoplastic Agents; Architecture; Automobile Driving; Biocompatible Materials; Biological; Biomedical Engineering; Bone Marrow; Cancer Etiology; Cause of Death; Cell Membrane Permeability; Cell Proliferation; Cell secretion; Cells; Cessation of life; Circulation; Clinical; Collagen; Collagen Fiber; Colorectal Cancer; Colorectal Neoplasms; Development; Devices; Diagnosis; Disease; Distant; Drug Targeting; Engraftment; Environment; Event; Extracellular Matrix; Extravasation; Fibronectins; Fire - disasters; Future; Goals; Growth; Hematopoietic stem cells; Hepatic; Hepatocyte; Homing; Human; Hydrogels; Immune; Immune system; In Vitro; Inflammatory; Interruption; Intervention; Invaded; Kinetics; Kupffer Cells; Laminin; Lead; Link; Liquid substance; Liver; Measures; Mediator; Metastatic Neoplasm to the Liver; Methodology; Microfluidic Microchips; Microfluidics; Modeling; Motivation; Neoplasm Metastasis; Organ; Organoids; Paper; Patients; Phenotype; Population; Primary Neoplasm; Process; Proliferating; Publishing; Resources; Signal Transduction; Site; Stromal Cells; Targeted Research; Technology; Testing; Therapeutic Intervention; Tissue Engineering; Tissue Model; Tissues; Tumor Cell Line; cancer invasiveness; cancer therapy; cell motility; 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; predictive marker; predictive tools; prevent; recruit; risk prediction; stellate cell; time interval; tool; transcriptome sequencing; treatment response; tumor; tumor microenvironment

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 转移的主要部位；而且，大多数被诊断为CRC死亡的患者因肝而死 转移。这为研究复杂转移性背后的机制提供了强大的动力 级联，希望导致新颖的干预方法。但是，研究生物学机制 这导致转移性疾病一直很困难。最近出现的研究目标是 形成远离原发性肿瘤的远处的thementantic利基市场（PMN），但创造了有利的环境 用于转移的细胞占据。研究PMN是具有挑战性的，因为在动物模型中几乎不可能 为了预测PMN中何时发生变化，并在鉴定转移后，为时已晚。一个 临床上与临床上的体外模型，其中转移和导致转移的组织的变化可能是 观察和操纵将是探索这些机制并识别的宝贵资源 绕过它们的方法。我们的团队现在发表了几篇论文，描述了我们的生物工程体外 由微流体设备和3D细胞外基质（ECM）组成的转移片（MOC）平台（MOC）平台 基于水凝胶的肿瘤和组织器官。我们的目标是使用这些工具来身份并早日查明 由于原发性肿瘤分泌介质而发生的肝脏PMN的变化并评估其贡献 到CRC转移。我们将使用3D肝，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，以量化肝脏重塑和BM- HPC招募，然后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，肿瘤细胞系的分泌介质 类器官和患者衍生的肿瘤器官（PTO）的不同等级将诱导相应的光谱 ECM重塑，建筑失调和BM-HPC募集的招募，当增强时， 随着转移的增加。在AIM 1中，我们将对原发性人肝和BM-HPC器官进行CRC 有条件的培养基，测量肝基质（星状）和免疫（kupffer）细胞激活，并量化ECM 根据组成，体系结构和机械性能进行重塑。作为此目标的一部分，我们将剖析 排除星状细胞，kupffer细胞和BM-HPC对这些变化的相对贡献。在AIM 2中，我们的MOC 设备还将包括肿瘤类器官，此外，肝脏和BM-HPC器官都将评估PMN 在这个动态的多组织平台中形成。将测量从肿瘤到肝癌的转移。 这将使我们能够将PMN的更改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的不同特征的机会 在无法治愈之前，还可以作为早期转移过程的未来目标以及生物标志物 患者转移的预测。