A microfluidic blood-based test to monitor treatment response in glioblastoma

项目总结 胶质母细胞瘤患者的诊断和监测需要手术和磁共振检查。 在补体中成像。外科手术为组织学和分子定征和核磁共振成像提供了关键的组织 促进后续监测，尽管其限制区分治疗反应(辐射 失败(肿瘤复发)。因此，患者需要接受额外的外科手术才能获得 组织的分子特征，并区分治疗反应和失败需要改变 心理治疗。考虑到手术相关的发病率，患者不能接受连续活检，因此 微创检测将提供实时分析，并改变我们对 胶质母细胞瘤。我们的实验室是第一个鉴定出循环中的肿瘤细胞(CTCs)和外周小体(EVS)的 胶质母细胞瘤患者的血液。在其他癌症中，CTC和EV已经被独立捕获， 表征和分析，以提供患者肿瘤负担的实时信息，并识别新的 对治疗产生抵抗力的突变。尽管有血脑屏障，但在血液中发现了EVS和CTC 在高频和低频下。目前，不存在同时捕获和监控两者的现有技术 患者血液中的EVS和CTC。因此，我们建议使用微流控技术来开发和验证我们的血液 从胶质母细胞瘤患者的单管血液中分离EVS和CTC 在诊断和整个治疗过程中。我们还将使用标准技术来并行评估ctDNA 评估对我们化验结果的任何附加益处。临床样品的微流控加工成本低，显示效果好。 承诺将“芯片上的血液”分析转化为临床。我们的化验将允许实时分子 在整个治疗过程中进行特征描述，并可能确定新的治疗方法或更好地将患者与 现有的临床试验。此外，我们的目标是更准确地确定治疗反应和不可避免的 从诊断开始使用下一代CTCs、EVS和CTDNA测序进行复发；最终 建立一种诊断、监测和检测胶质母细胞瘤患者复发的微创检测方法。 2016年世界卫生组织对脑肿瘤的新分类增加了分子标志物用于脑肿瘤的病理诊断 胶质母细胞瘤，传统上需要手术组织；我们的目标是通过液体活检提供第一个 通过简单的血液测试实时对脑肿瘤进行分子表征和分类。这将是 极大地促进了我们对胶质母细胞瘤患者的理解和护理。

PROJECT SUMMARY The diagnosis and monitoring of patients with glioblastoma requires both surgery and magnetic resonance imaging in complement. Surgery critically provides tissue for histological and molecular characterization and MRI facilitates subsequent monitoring despite its limitation to differentiate between treatment response (radiation necrosis) and failure (tumor recurrence). As a result, patients undergo an additional surgical procedure to obtain tissue for molecular characterization, and to distinguish treatment response from failure requiring a change in therapy. Given the morbidity associated with surgery, patients cannot undergo serial biopsies therefore a minimally invasive test would provide real-time analysis and change our understanding and management of glioblastoma. Our laboratory is the first to identify both circulating tumor cells (CTCs) and exosomes (EVs) in the blood of patients with glioblastoma. In other cancers, CTCs and EVs have been independently captured, characterized, and analyzed to provide real-time information of the patient's tumor burden and identifying new mutations that confer resistance to therapy. Despite the blood brain barrier, EVs and CTCs are found in the blood in high and low frequencies. Currently, no existing technology exists to simultaneously capture and monitor both EVs and CTCs in patient blood. Thus, we propose the use of microfluidics to develop and validate our blood based `liquid biopsy' to isolate both EVs and CTCs from a single tube of blood from patients with glioblastoma at diagnosis and throughout treatment. We will also use standard techniques to evaluate ctDNA in parallel to evaluate any additive benefit to our assay. Microfluidic processing of clinical samples is low cost and shows great promise for translating `blood-on-a-chip' assays to the clinic. Our assay will allow for real-time molecular characterization throughout therapy and potentially identify novel treatments or better match patients with existing clinical trials. In addition, we aim to more accurately determine treatment response and inevitable recurrence using next generation sequencing of CTCs, EVs and ctDNA from diagnosis onward; ultimately establishing a minimally invasive test to diagnose, monitor, and detect recurrence in patients with glioblastoma. The new 2016 WHO classification of brain tumors has added molecular markers to the pathological diagnosis of glioblastoma which has traditionally required surgical tissue; we aim through our liquid biopsy to provide the first molecular characterization and classification of brain tumors through a simple blood test in real-time. This will dramatically advance our understanding and the care of patients with glioblastoma.