BRAVE hydrogels for interrogating cell-matrix interactions in pancreatic desmoplasia

项目总结/摘要 胰腺癌的5年生存率只有8%，预计将成为第二大病因。 到2030年，所有癌症相关死亡人数。虽然已经确定了许多针对胰腺癌的治疗方法， 这种疾病的治疗仍然是具有挑战性的，由于异常致密和血管化不足的基质， 组织.因此，更深入地理解粘弹性基质中的肿瘤-基质相互作用将有助于研究肿瘤-基质的相互作用。 鉴定对抗这种致命疾病的新分子靶点。聚合物水凝胶能够 细胞外基质（ECM）的重演方面是研究癌细胞命运的理想选择，因为这些凝胶可以 可以被工程化为具有精确的生物物理和/或生物化学性质，以模拟肿瘤ECM。很长的- 本项目的长期目标是使用生物启发，响应和粘弹性（BRAVE）矩阵来阐明 控制胰腺癌细胞（PCCs）进展的分子机制，以及用于鉴定新的 分子靶点，以改善治疗效果。在R 01项目中，我们将开发BRAVE水凝胶 由功能化的透明质酸和明胶组成，还含有胶原蛋白和纤维连接蛋白， 指导胰腺细胞行为的基质因子。在目标1中，我们将开发所述BRAVE水凝胶以定义 主要基质组分之间的协同作用（例如，HA、FN和基质粘弹性）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测定（HCA）相容的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 共培养PCC和患者来源的癌症相关成纤维细胞（PD-CAF）的机制， 基质细胞这个平台将提供直接的实验证据有关的必要性，加强矩阵 关于PCC中的细胞间串扰和EMT。在目标3中，我们将开发具有梯度刚度的BRAVE凝胶， PCC迁移（即，硬旋转）在各种基质因素的影响下（例如，细胞因子，CAF，和 抑制剂）。这些结果将提供有关PCC迁移/侵袭和潜在分子靶点的见解 抗PCC转移。本提案中开发的BRAVE水凝胶和硬组织扩张装置将使我们能够 回答许多胰腺癌相关的问题，否则很难解决。而且 该项目的成果将对其他癌症的基础和应用研究产生影响， 用于组织再生的干细胞分化。

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.