Supramolecular matrix materials for prostate cancer cell biology

前列腺癌的发病率和死亡率仍然是重要的临床问题， 新的治疗策略取决于提高我们对前列腺癌生物学的基本理解。等 基础研究反过来又依赖于准确和相关的实验模型，无论是在体外， in vivo.然而，基础前列腺癌研究受到了建立患者- 衍生的细胞系和充分定制的3D细胞培养基质的不可用性。目前 大多数人前列腺癌细胞系来源于去势抵抗性转移瘤， 与其他癌症如胰腺癌相比，来自临床的高级别前列腺肿瘤， 标本不能有效地在动物模型中作为异种移植物建立。当前3D细胞培养基质 例如Matrigel已经使得该领域能够建立一些前列腺癌类器官培养物，但是Matrigel的 其有用性受限于其差的批次间一致性和难以调整其组成， 特定的细胞类型和特定的环境。因此，开发更明确和可定制的矩阵 将克服建立和维持前列腺癌细胞类器官的主要技术障碍， 基础研究。Collier实验室最近开发了一种策略，用于从定义的一组凝胶材料中创建凝胶材料。 表达的蛋白质，使用分子自组装。在这种方法中，蛋白质被工程化以展示一种 一种新的标签，允许细菌以单体状态表达，但随后自组装成 在与短的合成肽混合后，确定了3D培养基质。这些材料可以配制成 到组成定义的细胞培养材料，使我们在拟议的工作，工程改进 支持前列腺癌类器官生长的3D培养基。这项工作将通过两个 目标如下。目的1：开发用于前列腺癌细胞体外生长的定制细胞培养基质， 使用新型自组装蛋白（β-Tails）和自组装肽在体内引发肿瘤。 目的2：开发前列腺特异性抗原（PSA）响应性3D培养基，并将其工程化， 通过在细胞分泌PSA时释放FGF 10加速原代人前列腺细胞生长。我们 期望这些可定制的细胞培养基质不仅可用于培养患者来源的 前列腺癌细胞，而且还用于各种其他细胞类型。这个项目将是一个关键的 概念验证将为这些材料的未来发展奠定基础， 不同的文化背景。两名PI，Collier博士和Vander Griend博士，是在同一时间任命的。 部门（外科），促进这种合作，旨在解决长期存在的问题，不适合 用于前列腺类器官培养的基质。

Prostate cancer incidence and mortality continue to be significant clinical problems, and the development of new treatment strategies depends on improving our basic understanding of prostate cancer biology. Such basic investigations in turn are dependent on accurate and relevant experimental models, both in vitro and in vivo. However, basic prostate cancer research has been challenged by difficulties in establishing patient- derived cell lines and the unavailability of adequately tailored 3D cell culture matrices. Currently, the majority of human prostate cancer cell lines are derived from castration-resistant metastases, and in contrast to other cancers such as pancreatic cancer, high-grade prostate tumors derived from clinical specimens do not efficiently establish in animal models as xenografts. Current 3D cell culture matrices such as Matrigel have enabled the field to establish some prostate cancer organoid cultures, but Matrigel’s usefulness is limited by its poor batch-to-batch consistency and difficulties in adjusting its composition for specific cell types and specific contexts. Thus, the development of more defined and customizable matrices would overcome a major technical hurdle in establishing and maintaining prostate cancer cell organoids for basic study. The Collier lab has recently developed a strategy for creating gel materials from defined sets of expressed proteins, using molecular self-assembly. In this approach, proteins are engineered to display a novel tag that allows bacteriologic expression in a monomeric state, but subsequent self-assembly into defined 3D culture matrices upon mixing with short synthetic peptides. These materials can be formulated into compositionally defined cell culture materials, allowing us in the proposed work to engineer improved 3D culture media that support prostate cancer organoid growth. The work will be conducted through two aims, as follows. Aim 1: Develop a tailored cell culture matrix for prostate cancer cell growth in vitro and tumor initiation in vivo, using novel self-assembling proteins (beta-Tails) and self-assembling peptides. Aim 2: Develop prostate-specific antigen (PSA)-responsive 3D culture media and engineer them to accelerate primary human prostate cell growth by releasing FGF10 as the cells secrete PSA. We expect these customizable cell culture matrices to be useful not only for culturing patient-derived prostate cancer cells, but also for a wide variety of additional cell types. This project will be a critical proof-of-concept that will establish the foundation of these materials for future development in a range of different culture contexts. The two PIs, Drs. Collier and Vander Griend, are appointed in the same department (Surgery), facilitating this collaboration aimed at solving the longstanding problem of unsuitable matrices for prostate organoid culture.