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: 10373531
• 负责人: Esak Lee
• 金额: $ 18.32万
• 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 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373531
• 关键词: 3-Dimensional; Affect; Animals; Biological; Biology; Biomedical Engineering; Biomimetics; Blood; Blood Circulation; Blood Vessels; Cadherins; Cancer Etiology; Carcinoma; Cell Line; Cells; Cessation of life; Clinical; Coculture Techniques; Collaborations; Dextrans; Diagnosis; Diffuse; Disease; Distant; Distant Metastasis; Endothelial Cells; Endothelium; Engineering; Epithelial; 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 of activation protein; 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 stage; Xenograft procedure; clinically relevant; clinically significant; improved; in vivo; in vivo Model; lymphatic vessel; 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 具有高度侵袭性，并在肿瘤进展的早期阶段在远处器官中形成转移。为了更好 为了了解 PDAC 转移，需要进一步评估肿瘤与血管的相互作用，因为肿瘤细胞 主要通过血液循环传播。然而，PDAC 如何与血管相互作用并建立 对远处转移知之甚少。最近，我们的研究使用三维（3D）仿生技术 PDAC 芯片和多个体内小鼠模型表明 PDAC 细胞侵入血管并 通过 ALK7 信号传导主动替换内皮细胞，导致肿瘤-血管混合结构的形成 PDAC 肿瘤。我们将这种现象称为肿瘤血管替代。尽管这一发现很新颖， PDAC 中肿瘤血管置换的生物学后果尚不清楚。了解 肿瘤血管置换的表型后果对于确定临床相关性和 阻断 PDAC 中 ALK7 的意义。我们假设 PDAC 肿瘤血管置换会增加肿瘤 血管渗透性；然后通过促进肿瘤细胞进入肿瘤细胞来促进肿瘤的浸润和转移。 血液循环通过渗漏的血管。为了检验这些假设，我们的目标是确定——在 体外和体内——(i) 如果肿瘤血管置换引起肿瘤血管渗漏并促进转移 (ii) ALK7 抑制或 ALK7 敲除 (KO) 是否改善或逆转肿瘤血管渗漏 和转移。在目标 1 中，我们将评估 ALK7 在周细胞覆盖的血液中 PDAC 血管通透性中的作用 通过共培养微血管内皮细胞和周细胞来模拟生理血管，从而实现芯片上血管 被周细胞包围（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 通过静脉注射葡聚糖分子来测量血管通透性（目标 1.2）。在目标 2 中，我们将检查 ALK7 通过在储库中建立转移前肝脏微环境来进行体外 PDAC 转移 连接到工程血管。将评估多个 PDAC 系以测试 ALK7 是否介导 肿瘤血管置换影响转移扩散（目标 2.1）。然后我们将评估 ALK7 在 PDAC 中的作用 与 Manuel Hidalgo 博士合作，使用人类患者来源的异种移植 (PDX) 模型进行体内转移。 将评估对照组与 ALK7 KO 组的转移性肿瘤负荷，以及循环肿瘤的数量 将确定细胞和总体存活率（目标 2.2）。总之，我们的 3D PDAC 片上系统将提供 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 破译 ALK7 信号传导在介导肿瘤血管失调和转移中的作用；并评估 我们是否能够通过靶向 ALK7 来减少 PDAC 进展和转移。