Adaptable hydrogel platform to study pancreatic cancer

描述（由申请人提供）：胰腺癌是所有癌症相关死亡的第四大原因。众所周知，它很难诊断，治疗选择非常有限。胰腺导管腺癌（PDAC）占所有形式胰腺癌的80%以上。了解PDAC细胞的生长和运动性是开发治疗PDAC的有效药物疗法的关键。迄今为止，大多数体外PDAC研究都是在非自然刚性（E > 1 GPa）的二维（2D）组织培养塑料皿（TCP）上进行的。2D表面的弹性刚度不仅使附着的细胞不自然地极化，而且由于异常的机械感测而导致细胞表现不同。2D细胞培养技术在肿瘤细胞的研究中也是不充分的，肿瘤细胞高度受基质组织的影响（即，结缔组织增生，由活化的胰腺星状细胞沉积的致密细胞外基质）。还认为结缔组织增生与胰腺癌细胞沟通以促进肿瘤进展并阻碍药物渗透和功效。尽管一些研究已经探索了3D基质（如Matrigel）在PDAC研究中的应用，但市售基质机械性能较弱，并且含有不明确和不可控的生化组分，这可能混淆实验结果。我们的中心假设是，一个合成的肿瘤生态位与动态和模块化的适应性，可用于阐明细胞外基质（ECM）线索的影响，PDAC细胞的生长，形态发生和药物疗效。为此，我们将开发合成水凝胶，其可以通过与PDAC相关的范围内的细胞相容性光暴露（365- 430 nm）可逆地固化/软化。在目标1中，我们将开发创新的动态促结缔组织增生模拟水凝胶基质以包封PDAC细胞（例如，PANC-1、科洛-357和ASPC 1），并研究它们的生长、形态发生和上皮-间充质转化（EMT）。在目标2中，我们将独立地和可逆地调节生物物理和生物化学性质的细胞负载的水凝胶，以描绘各种固定和可溶性细胞外因子对PDAC细胞命运过程的影响。我们还将揭示这些关键因素对PDAC耐药性的影响。从这项研究中获得的信息将为治疗致死性PDAC开辟新的治疗选择。

DESCRIPTION (provided by applicant): Pancreatic cancer is the fourth leading cause of all cancer-related death. It is notoriously difficult to diagnose and has very limited therapeutic options. Pancreatic ductal adenocarcinoma (PDAC) accounts for more than 80% of all forms of pancreatic cancer. Understanding PDAC cell growth and motility is the key to developing effective drug therapeutics for treating PDAC. To date, most in vitro PDAC studies have been conducted on two dimensional (2D) tissue-culture plastic dishes (TCP) that are unnaturally stiff (E > 1 GPa). The ultrahigh stiffness of a 2D surface not only un-naturally polarizes the attached cells, but also causes the cells to behave differently due to abnormal mechano-sensing. 2D cell culture techniques are also inadequate in the study of tumor cells, which are highly influenced by the stromal tissues (i.e., desmoplasia, dense extracellular matrices deposited by activated pancreatic stellate cells) found in three-dimensional (3D) tumors. It is also believed that desmoplasia communicates with pancreatic cancer cells to promote tumor progression and to hinder drug penetration and efficacy. Although a few studies have explored the utility of 3D matrices, such as Matrigel(R), for PDAC research, the commercially available matrices are mechanically weak and contain ill-defined and un- controllable biochemical components that may confound the experimental results. Our central hypothesis is that a synthetic tumor niche with dynamically and modularly adaptable properties can be used to elucidate the influence of extracellular matrix (ECM) cues on the growth, morphogenesis, and drug efficacy in PDAC cells. Toward this end, we will develop synthetic hydrogels that can be reversibly stiffen/soften via cytocompatible light exposure (365-430nm) in a range relevant to PDAC. In Aim 1, we will develop innovative dynamic desmoplasia-mimetic hydrogel matrix to encapsulate PDAC cells (e.g., PANC-1, COLO-357, and ASPC1) and study their growth, morphogenesis, and epithelial-mesenchymal transition (EMT). In Aim 2, we will independently and reversibly modulate the biophysical and biochemical properties of cell-laden hydrogels in order to delineate the influence of various immobilized and soluble extracellular factors on PDAC cell fate processes. We will also reveal the influence of these critical factors on drug resistance in PDAC. The information obtained from this study will open new therapeutic options for treating the lethal PDAC.