Microfluidic Device to Profile Chemosensitivity in Glioma Slice Cultures

描述：目标：我们建议开发一个治疗性筛选平台，该平台可以快速确定完整的人类肿瘤样本对多种潜在疗法的平行反应。其目的是在足够快的时间范围内确定对个别患者具有最大潜在价值的治疗方法的子集，以指导治疗决策。创新：我们开发了一种微流体灌流系统，该系统可以在培养中保持原发肿瘤切片组织的完整，并能够将大量不同的药物、药物组合或药物方案阵列输送到肿瘤的解剖定义区域。我们的系统改进了现有的化疗药物活性筛选模型(例如，培养中的肿瘤细胞、小鼠异种移植或基因工程小鼠模型)：它使用保留人类肿瘤微环境的完整肿瘤样本，并允许在时间框架内生成响应数据，以指导治疗初始阶段的决策。焦点/目标：我们建议的研究重点是人脑肿瘤，特别是占原发成人脑肿瘤的大多数(70%)的恶性胶质瘤。这些肿瘤包括III级恶性胶质瘤和更常见的IV级多形性胶质母细胞瘤(GBM)。尽管几十年来一直在努力改善治疗结果，但被诊断为GBM的患者的中位生存期为~1年，总的5年生存率为5%。在这项提案中，我们认为，在治疗这些致命的、难治性肿瘤方面取得进展的一个重要方法是开发一种方法，在个体患者完整的GBM标本上快速测试许多潜在的治疗药物或方案。肿瘤对药物的反应将通过细胞死亡和细胞存活的标志物成像来衡量。由于单个肿瘤样本往往是异质的，因此我们的微流控系统能够在样本的不同区域进行多路药物测试是至关重要的。阳性药物反应在整个肿瘤中应该是一致的和特异的。这个多元化的平台还将使研究可能对治疗有用的特定药物组合和剂量成为可能。影响：这项工作解决了开发方法和设备的迫切需要，以快速和可靠地评估通常不同的、完整的原发肿瘤标本对一系列可能的治疗方法的反应，目的是确定对个别肿瘤和患者最有效的治疗子集。

DESCRIPTION: Goal: We propose to develop a therapeutic screening platform that rapidly determines the response of intact human tumor samples to many potential therapies in parallel. The aim is to identify the subset of therapies of greatest potential value to individual patients, n a timescale rapid enough to guide therapeutic decision- making. Innovation: We have developed a microfluidic perfusion system that maintains intact primary tumor slice tissue in culture and enables the arrayed delivery of large numbers of different drugs, drug combinations or drug regimens to anatomically-defined regions of the tumor. Our system improves upon existing models for screening chemotherapeutic drug activity (e.g. tumor cells in culture, mouse xenografts, or genetically engineered mouse models): it uses intact tumor samples that retain the human tumor microenvironment and allows the generation of response data in a time frame that can guide decision-making for the initial phases of therapy. Focus/Aims: Our proposed research focuses on human brain tumors, specifically the malignant gliomas that comprise the majority (70%) of primary adult brain tumors. These include Grade III malignant gliomas and the more common Grade IV glioblastoma multiforme (GBM). Patients diagnosed with GBM have a median survival of ~1 year and an overall 5-year survival of <5 %, despite decades of effort to improve treated outcomes. In this proposal we argue that an important way to make progress in treating these lethal, refractory tumors is to develop a way to rapidly test many potential therapeutic agents or regimens in parallel on intact GBM specimens from individual patients. Tumor response to drugs will be measured by imaging of markers for cell death and for cell viability. As individual tumor samples are often heterogeneous, it is critical that our microfluidi system enables multiplexed drug testing across different regions of the sample. Positive drug responses should be consistent and specific across the tumor. The multiplexed platform will also enable investigation of specific drug combinations and dosages that might be therapeutically useful. Impact: This work addresses the urgent need to develop approaches and devices to rapidly and reliably assess the response of often heterogeneous, intact primary tumor specimens to a range of possible therapies, with the goal of identifying the most effective subset of therapies for individual tumors and patients.