High-Throughput Ex Vivo Tumor Cell Motility Assay

描述(由申请人提供)：胶质瘤是最具侵袭性但治疗最不成功的脑肿瘤，因为它们具有侵袭周围脑组织的独特能力。即使大部分肿瘤被切除，留下的迁移细胞也不会被免疫系统检测到，并抵抗目前的细胞毒疗法。然后，肿瘤复发通常会导致迅速而致命的后果。了解胶质瘤细胞迁移的机制和设计新的靶向策略是设计更成功的治疗方法的主要挑战。 使用高度协作的多学科方法，我们将研究引导胶质瘤在生物兼容纳米纤维上迁移的微环境和地形线索，这些纳米纤维旨在模拟大脑内的底物地形。我们的具体目标是：1)优化高通量的体外迁移实验，使用模拟纳米级神经组织形貌的电纺纤维来确定该模型作为体外临床脑胶质瘤样本运动性预测生物实验的潜力；2)利用高通量实验来探索临床胶质瘤细胞直接迁移的机制和调节运动性的分子因素；以及3)分析针对细胞扩散的抗迁移策略，以确定该模型作为临床脑胶质瘤样本运动性预测生物实验的潜力。为了实现这些目标，我们将1)生产底部具有排列的、电纺的纳米纤维的高通量多孔细胞培养板，以模拟白质的排列结构；2)分析胶质瘤细胞的迁移并表征在呈现不同形貌的纳米纤维上发生的细胞/分子变化；3)在这些设备上研究潜在的分子靶点和抗迁移化疗药物；4)分析肿瘤细胞的体外迁移并在这些样本上测试抗迁移策略，以确定纳米纤维模型作为具有直接临床意义的预测生物检测的潜力。 这些目标涉及加速创新研究小企业催化剂奖(R43)的目标和范围，并与NIH的健康使命直接相关，同时显示出巨大的商业潜力。我们预计这些研究将提供更好、更准确的胶质瘤转移模型，具有更好的预测能力和更高的翻译潜力，以改善公众健康。与此同时，我们相信，我们的研究可能会确定调控胶质瘤细胞迁移的关键因素，可能有助于设计出一系列针对这些毁灭性肿瘤的有效治疗方法。如果这种高通量运动性分析在这项工作中被证明是成功的，那么它将为来自胶质瘤以外各种背景的研究人员提供一个创新的工具。 公共卫生相关性：恶性脑瘤具有高度侵袭性，可导致周围正常脑组织的浸润。这使得即使在手术切除了主要肿瘤之后，它们也极难或不可能被“治愈”。如果要有效地管理这些疾病，针对这些迁移细胞群体的治疗目标被认为是至关重要的。为了确定改进的治疗方法，我们提出了一个创新的模型，使用生物兼容的纳米纤维来模拟调节胶质瘤细胞迁移的大脑结构的局部方面。我们将使用这一新的模型来检测临床样本并预测预防肿瘤复发的策略。如果证明成功，我们建议这种创新的高通量设备将具有巨大的商业价值，因为它将有助于设计出新的方法，从而改善患者的结果。

DESCRIPTION (provided by applicant): Gliomas are the most aggressive and least successfully treated brain tumors because of their distinctive ability to infiltrate surrounding brain tissue. Even if the bulk of the tumor is removed, migratory cells left behind are not detected by the immune system and resist current cytotoxic therapies. Tumor recurrence then typically involves a rapid and deadly outcome. Understanding the mechanisms of glioma cell migration and designing novel targeting strategies are major challenges in devising more successful therapies against these tumors. Using a highly collaborative, multidisciplinary approach, we will investigate microenvironmental and topographic cues that direct glioma migration on biocompatible nanofibers designed to mimic the substrate topography within the brain. Our specific aims are: 1) optimize a high-throughput in vitro migration assay using electrospun fiber mimicking nanoscale neural tissue topography to determine the potential of this model as a predictive bioassay of motility in ex vivo clinical glioma samples; 2) utilize the high-throughput assay to explore mechanisms that direct migration of clinical glioma cells and molecular factors regulating motility; and 3) assay anti-migratory strategies against cell dispersion to determine the potential of this model as a predictive bioassay of motility in clinical glioma samples. To pursue these aims we will 1) produce high-throughput multiwell cell culture plates with aligned, electrospun nanofiber on the bottom to mimic the aligned structure of white matter, 2) analyze glioma cell migration and characterize the cellular/molecular changes that occur on nanofibers presenting different topographies, 3) investigate potential molecular targets and anti-migratory chemotherapeutics against glioma migration on these devices, and 4) analyze the migration of ex vivo tumor cells and test anti-migratory strategies on those samples to determine the potential of the nanofiber model as a predictive bioassay with immediate clinical relevance. These aims address the goals and scope of the Small Business Catalyst Awards for Accelerating Innovative Research (R43) and are directly relevant to the health mission of the NIH while showing a large commercial potential. We expect that these studies will provide improved, more accurate models of glioma migration, having better predictive power and higher translational potential to improve public health. At the same time, we believe that our research may identify key factors that regulate glioma cell migration, potentially helping to devise a broad range of effective therapies against these devastating tumors. If this high-throughput motility assay is proven successful in this work, then it will provide an innovative tool to researchers from a large variety of backgrounds beyond gliomas. PUBLIC HEALTH RELEVANCE: Malignant brain tumors are highly invasive leading to infiltration of the surrounding normal brain tissue. This makes them extremely difficult or impossible to 'cure' even after surgical removal of the main tumor. Therapeutic targeting of this migrating cell population is thought to be essential if these diseases are to be managed effectively. In order to identify improved therapeutic approaches, we propose an innovative model using biocompatible nanofibers to mimic local aspects of brain structure that regulate glioma cell migration. We will use this novel model to assay clinical samples and predict strategies to prevent tumor recurrence. If proven successful, we propose that this innovative high-throughput device will have a large commercial value as it will help devise novel approaches that lead to improved patient outcomes.