Targeting the Plasminogen Activation System to Limit Pancreatic Cancer Progression and Associated Thrombosis

摘要 胰腺导管腺癌(PDAC)是所有癌症中死亡率最高的癌症之一， 血栓栓塞症并发症发生率最高。PDAC的血栓前潜能直接与 PDAC肿瘤细胞高水平表达组织因子促进凝血酶活性增强 在血液循环和肿瘤微环境中。这项建议是基于侵袭性肿瘤 PDAC的进展与血栓形成通过关键的双向机制联系在一起 肿瘤生长和促凝血潜能。初步数据显示，高水平的肿瘤细胞转铁蛋白活性领先于 到PDAC肿瘤细胞和肿瘤相关成纤维细胞(CAF)的凝血酶/PAR-1信号转导，驱动局部 生产纤溶酶原激活(PA)系统组件、尿激酶型纤溶酶原激活剂(UPA)和 受体(UPAR)。初步结果还表明，PA系统是推动双向 PDAC肿瘤进展和静脉血栓栓塞症并发症的机制。这些发现支持 以PA系统组件为目标将限制PDAC进展的中心假设 同时矛盾地抑制PDAC相关的血栓形成。拟议的研究将直接 明确PAR-1/uPA/uPAR轴与PDAC肿瘤生长和侵袭的前馈机制 并将确定uPA/uPAR/纤溶酶原在PDAC中驱动血栓形成的反馈机制。 该联盟是由基础和临床止血、肿瘤生物学、 和生物工程。拟议的研究将使用多种创新方法，包括分析 Novo PDAC在独特的转基因小鼠中，一种尖端的3D仿生培养系统，新颖 药理学工具和患者来源的翻译异种移植模型来分析人类肿瘤细胞和 CAF。这一强大的专业知识和试剂集合将被用来检验以下具体假设： (1)par-1驱动的uPA和uPAR在PDAC肿瘤中的表达促进肿瘤进展，并驱动 通过调节肿瘤细胞相关的TF促凝活性的释放来促进循环的血栓形成 促凝血活性；(2)肿瘤细胞固有的PAR-1活性通过诱导 UPA/uPAR，而CAF的PAR-1信号驱动uPA/uPAR介导的组织重塑 先进的PDAC；和(3)针对PA的单独或与抗凝联合治疗将 显著抑制PDAC肿瘤进展和相关的血栓形成。建议进行的研究 将为PAR-1/uPA/uPAR在PDAC病理生物学中的贡献提供新的见解，阐明关键 PA系统与PDAC相关性血栓形成的偶联机制，并提供必要的证据 实验动物的基本数据和患者衍生的材料，以促进将研究结果转化为 治疗PDAC和癌症相关血栓形成的新疗法。

SUMMARY Pancreatic ductal adenocarcinoma (PDAC) has one of the highest mortality rates of all cancers, and the highest rate of thromboembolic complications. The prothrombotic potential of PDAC has been directly linked to high-level expression of Tissue Factor (TF) by PDAC tumor cells that drives exuberant thrombin activity in circulation and in the tumor microenvironment. This proposal is based on evidence that aggressive tumor progression and thrombophilia in PDAC are integrally linked via key bi-directional mechanisms promoting tumor growth and procoagulant potential. Preliminary data suggests that high-level tumor cell TF activity leads to thrombin/PAR-1 signaling in PDAC tumor cells and cancer-associated fibroblasts (CAFs), driving local production of the plasminogen activation (PA) system components, urokinase plasminogen activator (uPA) and receptor (uPAR). Initial results also indicate that the PA system is a central player driving the bi-directional mechanisms of PDAC tumor progression and venous thromboembolic complications. These findings support the central hypothesis that targeting PA system components will limit PDAC progression as well as simultaneously and paradoxically suppress PDAC-associated thrombophilia. The proposed studies will directly define the feed forward mechanisms linking the PAR-1/uPA/uPAR axis to PDAC tumor growth and invasion and will also identify the feedback mechanisms by which uPA/uPAR/plasminogen drive thrombophilia in PDAC. This consortium is a collaboration of experts in the basic and clinical sciences of hemostasis, tumor biology, and bioengineering. The proposed studies will use multiple innovative approaches, including analysis of de novo PDAC in unique genetically-modified mice, a cutting-edge 3D biomimetic culture system, novel pharmacological tools, and translational patient-derived xenograft models to analyze human tumor cells and CAFs. This powerful collection of expertise and reagents will be used to test the following specific hypotheses: (1) PAR-1-driven expression of uPA and uPAR in PDAC tumors promotes cancer progression, and drives thrombosis by mediating the release of tumor-cell associated TF procoagulant activity that increases circulating procoagulant activity; (2) tumor cell-intrinsic PAR-1 activity supports PDAC invasion through induction of uPA/uPAR, while PAR-1 signaling by CAFs drives uPA/uPAR-mediated tissue remodeling associated with advanced PDAC; and (3) therapies targeting PA either alone or in combination with anticoagulation will significantly impede both PDAC tumor progression and the associated thrombophilia. The proposed studies will provide novel insights into the contribution of PAR-1/uPA/uPAR to PDAC pathobiology, illuminate key mechanisms coupling the PA system to PDAC-associated thrombophilia, and provide essential proof-of- principle data in experimental animals and with patient-derived material to facilitate translation of findings into new treatments for PDAC and cancer-associated thrombosis.