BRAVE hydrogels for interrogating cell-matrix interactions in pancreatic desmoplasia

项目摘要/摘要 胰腺癌的5年存活率低得可怜，只有8%，预计将成为第二大死因。 到2030年，所有与癌症相关的死亡人数。虽然已经确定了许多针对胰腺癌的治疗方法， 由于间质异常致密和少血管，这种疾病的治疗仍然具有挑战性。 组织。因此，更深入地了解粘弹性基质中肿瘤-间质的相互作用将有助于 针对这种致命疾病的新分子靶点的鉴定。聚合物水凝胶能够 概括地说，细胞外基质(Ecm)是研究癌细胞命运的理想材料，就像这些凝胶一样。 具有精确的生物物理和/或生化特性，模拟肿瘤细胞外基质。长的- 本项目的学期目标是使用生物启发、响应和粘弹性(VERE)矩阵来阐明 控制胰腺癌细胞(PCCs)进展的分子机制以及识别新的 改善治疗结果的分子靶点。在这个R01项目中，我们将开发勇敢的水凝胶 由功能化的透明质酸和明胶以及用于审讯的胶原和纤维连接蛋白组成 引导胰腺细胞行为的基质因子。在目标1中，我们将开发上述勇敢水凝胶，以定义 主要基质成分(如HA、FN和基质粘弹性)之间的协同作用对PCC进展的影响。 在目标2中，我们将制备与细胞命运过程的高含量分析(HCA)兼容的BRAVE水凝胶。 我们还将设计具有相同生化成分但空间分级的双层Brave凝胶 PCCs与患者肿瘤相关成纤维细胞(PD-CAF)共培养的机制 基质细胞。该平台将提供关于加强矩阵的必要性的直接实验证据 关于PCCs中的细胞-细胞串扰和EMT。在目标3中，我们将开发具有梯度硬度的勇敢凝胶来审问 在各种基质因素(例如，细胞因子、CAF和 抑制剂)。这些结果将提供关于PCC迁移/入侵和潜在分子靶点的见解 抗PCC转移。在这项提案中开发的勇敢水凝胶和趋杜性设备将使我们能够 回答许多与胰腺癌相关的问题，否则很难解决。此外， 该项目的成果将对其他癌症的基础和应用研究以及定向研究产生影响 干细胞分化促进组织再生。

PROJECT SUMMARY/ABSTRACT With a dismal 5-year survival rate of ~8%, pancreatic cancer is projected to become the second leading cause of all cancer-related deaths by 2030. While many therapeutics against pancreatic cancer have been identified, treatment of this disease remains challenging owing to the exceptionally dense and hypovascularized stromal tissue. Therefore, deeper understanding of tumor-stroma interactions in a viscoelastic matrix will facilitate the identification of novel molecular targets against this deadly disease. Polymeric hydrogels capable of recapitulating aspects of the extracellular matrix (ECM) are ideal for studying cancer cell fate as these gels can be engineered with precise biophysical and/or biochemical properties that emulate the tumor ECM. The long- term objective of this project is to use bio-inspired, responsive, and viscoelastic (BRAVE) matrices for elucidating molecular mechanisms governing progression of pancreatic cancer cells (PCCs), as well as for identifying novel molecular targets to improve treatment outcome. In this R01 project, we will develop BRAVE hydrogels composed of functionalized hyaluronic acid and gelatin, as well as collagen and fibronectin for interrogating matrix factors guiding pancreatic cell behaviors. In Aim 1, we will develop the said BRAVE hydrogels to define the synergisms between major matrix components (e.g., HA, FN, and matrix viscoelasticity) on PCC progression. In Aim 2, we will prepare BRAVE hydrogels compatible with high-content assay (HCA) of cell fate processes. We will also design dual-layer BRAVE gels with identical biochemical compositions but spatially graded mechanics for co-culturing PCCs and patient-derived cancer associated fibroblasts (PD-CAFs), the major tumor stromal cells. This platform will provide direct experimental evidence regarding the necessity of a stiffened matrix on cell-cell crosstalk and EMT in PCCs. In Aim 3, we will develop BRAVE gel with gradient stiffness to interrogate PCC migration (i.e., durotaxis) under the influence of various matrix factors (e.g., cytokines, CAFs, and inhibitors). The results will provide insights regarding PCC migration/invasion and potential molecular targets against PCC metastasis. The BRAVE hydrogels and durotaxis device developed in this proposal will allow us to answer many pancreatic cancer relevant questions that are otherwise difficult to address. Furthermore, the outcome of this project will have impact on basic and applied research in other cancers, as well as on directed stem cell differentiation for tissue regeneration.