A novel microfluidic device to predict brain cancer prognosis and response to therapy

项目摘要/摘要 脑瘤是导致儿童实体瘤死亡的首要原因，也是导致癌症的首要原因之一。 20-39岁青年男性和女性死亡 。胶质母细胞瘤是最常见的原发肿瘤。 成人脑癌的总发病率占所有原发脑瘤的20%。尽管目前的标准是 包括手术和化疗在内的治疗，中位生存期只有14.6个月。由于高度的 基底膜细胞的侵袭性，大体全切除肿瘤的标准治疗不足以 由于肿瘤切缘外的肿瘤细胞复发率高，达到治愈的目的。 此外，在临床医生识别存活时间较短的患者和 复发风险高。开发新的预后工具来确定患者的存活率和识别 细胞亚群的运动性和侵袭性显著增强，并驱动复发 非常重要。基于对我们的GBM患者的回顾研究的耐人寻味的数据显示：1)我们的新奇 微流体侵袭网络设备(Mind)基于增加的GbM患者的生存预测 异质种群中侵袭性细胞的百分比；2)我们能够将侵袭性细胞与 使用Mind设备的非侵入性细胞，这将使我们了解潜在的分子差异 3)靶向转录因子TEAD4选择性过表达 和过度活跃的基底膜，促进受限的基底膜迁移。根据这些调查结果，我们建议：i) 前瞻性地将我们的思维平台作为GBM患者临床管理的辅助工具，ii) 发现GBM细胞侵袭的新驱动因素，以及iii)将其用作高通量检测以筛选潜在的 治疗药物。为了实现这些目标，我们将追求以下具体目标：确定 在使用Mind的GBM患者的前瞻性队列中，受限空间迁移的预后能力(目标1)； 表征表型侵袭性脑癌细胞有限空间迁移的分子驱动因素 使用Mind(目标2)；评估42种化疗药物在体内的翻译效果 头脑(目标3)。体外实验将使用预期收集的患者来源的原代细胞进行 来自100多名患者的GBM细胞株。我们专有的、新颖的思维平台将用于研究受限的- 这些患者的空间迁移以确定预后不良和复发率高的前瞻性 未来的目标是实现患者的个性化治疗。体内研究将使用我们的 建立小鼠原位人脑肿瘤起始细胞来源的基底膜模型。从以下方面获得的结果： 这项研究将有助于开发一个具有在计算机上进行药物测试的精准医学平台 使用新鲜分离的人体组织的高通量方式。

PROJECT SUMMARY/ABSTRACT Brain tumor is the leading cause of solid tumor death in children and one of the top leading causes of cancer death in young male and female adults with ages of 20-39 . Glioblastoma (GBM) is the most common primary brain cancer in adults with an overall incidence of 20% of all primary brain tumors. Despite current standard of care, which includes surgery and chemo-radiation, the median survival is only 14.6 months. Due to the highly invasive nature of GBM cells, standard treatment with gross total resection of the tumor is insufficient to achieve a cure because of the high rate of recurrence by tumor cells outside the tumor resection margins. Moreover, there is an unmet need in the ability of clinicians to identify patients with lower survival times and high risk of recurrence. Developing novel prognostic tools for determining patient survival and for identifying cell subpopulations that have a significantly increased motility and aggressiveness, and drive recurrence is of utmost importance. Intriguing data based on a retrospective study of our GBM patients reveal that: 1) our novel Microfluidic Invasion Network Device (MIND) predicts survival in GBM patients based on the increased percentage of invasive cells in the heterogeneous population; 2) we are able to separate the invasive from non-invasive cell using MIND device, which will allow us to understand the underlying molecular differences of the more aggressive subpopulations; 3) the targetable transcription factor TEAD4 is selectively overexpressed and hyperactive in GBM and promotes confined GBM migration. In light of these findings, we propose to: i) establish our MIND platform as an adjunct tool to the clinical management of GBM patients prospectively, ii) uncover novel drivers of GBM cell invasion, and iii) utilize it as a high-throughput assay for screening potential therapeutic drugs. To achieve these goals, we will pursue the following specific aims: to determine the prognostic power of confined space migration in a prospective cohort of GBM patients using MIND (Aim 1); to characterize the molecular drivers of confined space migration of phenotypically aggressive brain cancer cells using MIND (Aim 2); to evaluate the in vivo translational efficacy of 42 chemotherapeutic agents tested in MIND (Aim 3). In vitro experiments will be conducted using prospectively collected patient-derived primary GBM cell lines from over 100 patients. Our proprietary and novel MIND platform will be used to study confined- space migration from these patients to identify poor prognosis and high recurrence prospectively with the future goal of achieving personalized treatments for patients. In vivo studies will be conducted using our established orthotopic human brain tumor initiating cell-derived GBM model in mice. The results obtained from this study will to lead to the development of a platform for precision medicine with the ability to test drugs in a high throughput fashion using freshly isolated human tissue.