Using photopolymerizable collagen matrices to investigate how progressive stiffening alters lymphatic trafficking of cancer and immune cells in pancreatic ductal adenocarcinoma

使用光聚合胶原基质研究渐进性硬化如何改变胰腺导管腺癌中癌症和免疫细胞的淋巴运输

• 批准号: 10359396
• 负责人: Catherine F Whittington
• 金额: $ 7.37万
• 依托单位: WORCESTER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359396
• 关键词: 3-Dimensional; Address; Adhesions; Adopted; Area; Attention; Biochemical; Biophysics; Cancer cell line; Cell Adhesion; Cell Communication; Cells; Chemotaxis; Collagen; Collagen Type I; Complex; Dendritic Cells; Dendritic cell tumor; Diagnosis; Disease; Disease Progression; Disease model; Drug Modelings; Endothelium; Environment; Equilibrium; Event; Exercise; Exposure to; Extracellular Matrix; Fibrosis; Foundations; Future; Goals; Growth; Heterogeneity; Hyaluronic Acid; Hydrogels; Immune; Immunosuppression; In Vitro; Individual; Knowledge; Light; Link; Lymphangiogenesis; Lymphatic; Lymphatic Capillaries; Lymphatic Endothelial Cells; Lymphatic Metastasis; Lymphocyte; Malignant Neoplasms; Methacrylates; Modeling; Neoplasm Metastasis; Normal tissue morphology; Pancreatic Ductal Adenocarcinoma; Pathology; Patient-Focused Outcomes; Patients; Phenotype; Pilot Projects; Population; Process; Research; Role; Specificity; Testing; Time; Tissue Engineering; Tumor Tissue; Tumor-associated macrophages; Tumor-infiltrating immune cells; Up-Regulation; Work; barrier to care; base; biophysical properties; cancer cell; cell behavior; cell motility; design; drug discovery; fluid flow; functional outcomes; improved; in vitro Model; in vivo; innovation; lymph nodes; lymphatic vasculature; lymphatic vessel; macrophage; mechanical force; migration; model design; monolayer; novel strategies; pancreatic ductal adenocarcinoma model; predictive modeling; response; shear stress; success; therapeutic target; therapy development; three-dimensional modeling; trafficking; tumor; tumor microenvironment; tumor-immune system interactions

PROJECT SUMMARY More effective in vitro models are needed to improve treatment for ~90% of patients with pancreatic ductal adenocarcinoma (PDAC) who die within five years of diagnosis. PDAC is deadly and difficult to treat due to high rates of metastasis and an immunosuppressive microenvironment. The disease progresses as lymphatic capillaries around the tumor transport metastasizing cancer cells and anti-tumor immune cells to the lymph nodes. PDAC also progresses, in part, through a fibrotic response (desmoplasia) that remodels the tumor extracellular matrix (ECM) and alters ECM stiffness over time. However, knowledge gaps remain on how the dynamic PDAC microenvironment regulates lymphatic interactions related to cancer and immune cell entry and trafficking, and most in vitro PDAC models lack temporal control over stiffness to mimic progressive ECM stiffening. Our objective is to use a PDAC-specific 3D in vitro model with progressive stiffening to investigate how changes in ECM stiffness alter early-stage trafficking events and immunosuppression. This pilot work contributes to the long-term goal of designing more complex, predictive models of PDAC progression for anti- tumor and anti-metastasis drug discovery and improved patient outcomes. Aims will test the central hypothesis that progressive ECM stiffening at the tumor periphery enhances cancer and immune cell trafficking via up- regulation of mechanosensitive molecules that enable migration before and after entering lymphatic capillaries. We propose two specific aims to accomplish our objective: (1) Define how progressive stiffening regulates lymphangiogenic, metastatic, immunosuppressive, phenotypes related to chemotaxis/migration, invasion, and maturation. Methacrylated type I collagen will be combined with hyaluronic acid to represent the PDAC ECM and allow the hydrogel to stiffen under light exposure to levels that represent normal and tumor tissues. Lymphatic endothelial cells, PDAC cancer cells, and immune cells (dendritic, macrophages) will be individually combined with hydrogels and assessed for phenotypic changes related to metastasis, trafficking, and immunosuppression. (2) Determine how mechanosensitivity regulates barrier integrity in lymphatic endothelial cells and adhesion and motility between lymphatic endothelial cells, PDAC cancer cells, and immune cells. Monolayers of lymphatic endothelial cells will be grown on hydrogels undergoing progressive stiffening. The monolayer will then be exposed to PDAC and immune cells that have also undergone stiffening to assess functional outcomes: adhesion strength, competitive adhesion, migration across the monolayer, endothelial barrier integrity, and transmural migration. This work is innovative in that it uses tissue engineering to exercise temporal control over ECM stiffness and center fibrosis and lymphatic interactions—key features that are missing from current in vitro PDAC models—in our understanding of PDAC progression. These pilot studies stand to impact PDAC treatment by making significant progress to investigate potential therapeutic targets of interactions that could that inhibit lymphatic metastasis and/or shift the balance of immunosuppressive and anti-tumor immune cells in PDAC.

项目摘要 需要更有效的体外模型来改善约90％的胰腺导管患者的治疗 诊断后五年内死亡的腺癌（PDAC）。 PDAC由于高而致命而难以治疗 转移率和免疫抑制微环境。该疾病作为淋巴发展 肿瘤周围的毛细血管转移转移的癌细胞和抗肿瘤免疫细胞向淋巴 节点。 PDAC还部分通过重塑肿瘤的纤维化反应（Desmoplasia）进行 细胞外基质（ECM）和随着时间的推移会改变ECM刚度。但是，知识差距仍然存在于如何 动态PDAC微环境调节与癌症和免疫细胞进入以及 贩运和大多数体外PDAC模型缺乏对模拟僵硬性ECM的刚度的暂时控制 僵硬。我们的目标是在体外模型中使用具有渐进式僵硬的PDAC特异性3D进行研究 ECM刚度的变化如何改变早期贩运事件和免疫抑制。这个飞行员工作 促进了设计更复杂，PDAC进展的更复杂，预测模型的长期目标 肿瘤和抗促药物发现并改善患者预后。目标将检验中心假设 肿瘤周围的渐进性ECM僵硬可通过上升来增强癌症和免疫细胞的运输 调节能够在进入淋巴毛细血管前后迁移的机械敏感分子的调节。 我们提出了两个具体目标以实现我们的目标：（1）定义渐进式僵硬的调节方式 淋巴血管，转移，免疫抑制作用，与​​趋化/迁移，侵袭和侵袭有关的表型 成熟。 I型I胶原蛋白的甲基丙烯酸将与氢酸合并，以代表PDAC ECM和 让水凝胶在暴露于代表正常组织和肿瘤组织的水平下变硬。淋巴 内皮细胞，PDAC癌细胞和免疫细胞（树突状，巨噬细胞）将单独组合 与水凝胶一起评估了与转移，运输和免疫抑制有关的表型变化。 （2）确定机理如何调节淋巴内皮细胞中的屏障完整性和粘合剂和 淋巴内皮细胞，PDAC癌细胞和免疫细胞之间的运动。淋巴管的单层 内皮细胞将在经历渐进式僵硬的水凝胶上生长。然后，单层将是 暴露于PDAC和免疫细胞也经历了僵硬以评估功能结果： 粘附强度，竞争性粘附，跨单层迁移，内皮屏障完整性和 透壁迁移。这项工作具有创新性，因为它使用组织工程进行临时控制 ECM刚度和中心纤维化和淋巴相互作用 - 当前体外缺少的关键特征 PDAC模型 - 在我们对PDAC进展的理解中。这些试点研究将影响PDAC治疗 通过取得重大进展来研究可能抑制的相互作用的潜在治疗靶标 淋巴转移和/或PDAC中免疫抑制和抗肿瘤免疫球的平衡。