A Bioengineered Model of Tumor Vessel Interactions in Pancreatic Cancer

胰腺癌肿瘤血管相互作用的生物工程模型

• 批准号: 10557226
• 负责人: Esak Lee
• 金额: $ 21.78万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557226
• 关键词: 3-Dimensional; Ablation; Affect; Animals; Biological; Biology; Biomedical Engineering; Biomimetics; Blood; Blood Circulation; Blood Vessels; Cadherins; Cancer Etiology; Carcinoma; Cell Line; Cells; Cellular Structures; Cessation of life; Circulation; Clinical; Coculture Techniques; Collaborations; Dextrans; Diagnosis; Disease; Distant; Distant Metastasis; Endothelial Cells; Endothelium; Engineering; Epithelium; Extravasation; Functional disorder; Goals; Hepatocyte; Human; Hybrids; Immune; In Vitro; Intravenous; Invaded; Knock-out; Liver; Lung; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Mediating; Mediator; Medicine; Mesenchymal; Metastatic Neoplasm to the Liver; Modeling; Mus; Names; Nature; Neoplasm Circulating Cells; Neoplasm Metastasis; Neoplasms in Vascular Tissue; Organ; Outcome; Pancreatic Ductal Adenocarcinoma; Patients; Pericytes; Permeability; Phenotype; Physiological; Process; Prognosis; Research Personnel; Role; Sampling; Signal Transduction; Stains; Stromal Cells; Stromal Neoplasm; Structure; Survival Rate; System; Testing; Therapeutic; Tissues; Tumor Burden; Tumor Cell Migration; Tumor Escape; Tumor Promotion; Tumor stage; Vascular Endothelial Cell; Vascularization; Xenograft procedure; clinically relevant; clinically significant; improved; in vivo; in vivo Model; lymphatic vessel; migration; mouse model; neoplastic cell; novel; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; patient derived xenograft model; prevent; stemness; tumor; tumor progression; tumor xenograft

Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer deaths among malignancies. PDAC is highly invasive and forms metastases in distant organs at the very early stage of tumor progression. To better understand PDAC metastasis, tumor-blood vessel interactions need to be evaluated further, as tumor cells spread primarily through the blood circulation. However, how PDAC interacts with blood vessels and establishes distant metastases are poorly understood. Recently, our study using both three-dimensional (3D) biomimetic PDAC-on-chip and multiple in vivo mouse models showed that PDAC cells invaded blood vasculatures and actively replaced endothelial cells via ALK7 signaling, leading to a formation of tumor-vessel hybrid structure in PDAC tumors. We refer to this phenomenon as tumor vessel replacement. Despite the novelty of the finding, it is unknown what the biological consequences of the tumor vessel replacement in PDAC are. Understanding the phenotypic consequences of the tumor vessel replacement is critical to determine the clinical relevance and significance of blocking ALK7 in PDAC. We hypothesize that PDAC tumor vessel replacement increases tumor vessel permeability; then promotes tumor intravasation and metastasis by facilitating tumor cells’ entering the blood circulation through the leakier vessels. In order to test these hypotheses, we aim to determine—in both in vitro and in vivo—(i) if tumor vessel replacement induces tumor vessel leakiness and promotes metastatic dissemination and (ii) if ALK7 inhibition or ALK7 knock out (KO) ameliorates or reverses tumor vessel leakiness and metastasis. In Aim 1, we will assess the role of ALK7 in PDAC vessel permeability in pericyte-covered blood vessel on-chip by co-culturing microvascular endothelial cells and pericytes to mimic physiological blood vessels surrounded by pericytes (Aim 1.1). Next, we will evaluate the role of ALK7 in PDAC vessel dysfunction in vivo. We will generate an orthotopic PDAC model using wild-type or ALK7-KO PDAC cells, and examine PDAC tumor vessel permeability by intravenously injecting dextran molecules (Aim 1.2). In Aim 2, we will examine ALK7 in PDAC metastasis in vitro by establishing pre-metastatic liver microenvironment in the reservoirs that are connected to the engineered blood vessel. Multiple PDAC lines will be assessed to test whether ALK7-mediated tumor vessel replacement affects metastatic spreading (Aim 2.1). We will then evaluate the role of ALK7 in PDAC metastasis in vivo using human patient-derived xenograft (PDX) models in collaboration with Dr. Manuel Hidalgo. Metastatic tumor burdens in control vs. ALK7 KO groups will be assessed, and the number of circulating tumor cells and overall survival rate will be determined (Aim 2.2). In summary, our 3D PDAC-on-chip system will provide a unique platform to better investigate PDAC interactions with blood vessels and metastatic progression. We will decipher the roles of ALK7 signaling in mediating tumor vessel dysregulation and metastasis; and assess whether we will be able to reduce PDAC progression and metastasis by targeting ALK7.

胰腺导管腺癌（PDAC）是恶性肿瘤中癌症死亡的主要原因。 PDAC是 在肿瘤进展的早期阶段，高度侵入性并形成远处器官的转移。更好 了解PDAC转移，需要进一步评估肿瘤血管相互作用，因为肿瘤细胞 主要通过血液循环传播。但是，PDAC如何与血管相互作用并建立 遥远的转移尚不理解。最近，我们使用三维（3D）仿生的研究 片上的PDAC和多个体内小鼠模型表明，PDAC细胞侵入血管和 通过ALK7信号传导积极地替代内皮细胞，导致形成肿瘤毒杂种结构 PDAC肿瘤。我们将这种现象称为肿瘤血管的替代。尽管这一发现有新奇 尚不清楚PDAC中肿瘤血管替代的生物学后果是什么。了解 肿瘤血管置换的表型后果对于确定临床相关性至关重要 阻塞ALK7在PDAC中的重要性。我们假设PDAC肿瘤血管置换会增加肿瘤 容器渗透性；然后通过支撑肿瘤细胞进入该肿瘤的侵入和转移 血液通过Leafier视频循环。为了检验这些假设，我们旨在确定 - 体外和体内 - （i）如果肿瘤血管置换会诱导肿瘤血管泄漏并促进转移性 传播和（ii）如果ALK7抑制或ALK7淘汰（KO）可以改善或逆转肿瘤容器的渗漏 和转移。在AIM 1中，我们将评估ALK7在PDAC血管渗透性中的作用 通过合作培养的微血管内皮细胞和周细胞的芯片容器，以模仿生理血管 被周围环绕（目标1.1）。接下来，我们将评估ALK7在体内PDAC血管功能障碍中的作用。 我们将使用野生型或ALK7-KO PDAC细胞生成原位PDAC模型，并检查PDAC肿瘤 通过静脉注射右旋葡萄糖分子（AIM 1.2），血管渗透性。在AIM 2中，我们将检查Alk7 通过在水库中建立肝脏微环境，在体外PDAC转移 连接到工程血管。将评估多个PDAC线以测试ALK7介导的 肿瘤血管替代会影响转移扩散（AIM 2.1）。然后，我们将评估ALK7在PDAC中的作用 使用人类患者衍生的Xenographotic（PDX）模型与Manuel Hidalgo博士合作，体内转移。 将评估对照与ALK7 KO组中的转移性肿瘤伯良，并评估循环肿瘤的数量 细胞和总生存率将确定（AIM 2.2）。总而言之，我们的3D PDAC片系统将提供 一个独特的平台，可以更好地研究PDAC与血管和转移进展的相互作用。我们将 解释了Alk7信号传导在介导肿瘤血管失调和转移中的作用；和评估 我们是否能够通过靶向ALK7来降低PDAC的进展和转移。