Imaging and Liquid Biopsy for Glioma Diagnosis and Treatment Monitoring

恶性胶质瘤(MG)是中枢神经系统最具侵袭性的恶性肿瘤，平均 存活时间为15个月。MG的快速进展和高度异质性要求频繁成像(MRI) 以测量治疗反应。然而，成像本身受到假阳性(“假进展”)的限制。 缺乏分子特异性。组织分析可以增强成像结果，但存在以下风险 共病和抽样误差。该项目的目标是应对这些挑战，并推动 微创，因此可连续重复的针对MG的临床检测。我们将开发细胞外 囊泡(EVS)作为一类新的循环肿瘤生物标志物。越来越多的研究证明电动汽车 潜在用途：这些囊泡从所有细胞中释放出来，作为可靠的细胞替代品发挥作用，并反映 全球肿瘤负担，克服了肿瘤异质性和抽样偏差的限制。我们现在寻求 建立一种用于MG诊断和治疗监测的翻译EV方法，并比较其性能 具有黄金标准的基于成像的诊断。首先，我们将标准化EV检测方案，用于临床 工作流程。我们将采用我们经过验证的技术：ExoLution，一种临床级别的EV分离试剂盒；Shahky，一种 用于EV蛋白分析的高通量等离子仪器；用于高灵敏度EV的数字液滴聚合酶链式反应 MRNA检测。利用Exosome诊断公司的开发和监管专业知识，我们将 使这些平台做好转化为临床诊断实验室的准备。第二，我们将执行一项 有针对性的临床研究，严格评估EVS对GBM的诊断和预后能力。我们会收集 GBM患者治疗前后循环EV及EV蛋白/mRNA动态变化监测 特别是用来检测获得性耐药性的早期迹象。我们将比较EV检测的结果 和伴随的MRI，并建立治疗监测的综合模型。我们组成了一个强大的 执行这些项目的多学科团队：马萨诸塞州总医院脑瘤中心 (MGH)，运行着一个庞大的人类临床样本生物库计划；系统生物学中心，位于 MGH是开发电动汽车分析新技术的先驱；Exosome诊断公司是事实上的 在基于EV的液体活检方面处于行业领先地位，在检测标准化方面拥有丰富的经验。总体而言， 该团队在研究EVS作为神经胶质瘤生物标记物的潜力方面有着良好的记录，并建立了最佳EV 用于分子分析的分析方法。我们将确保检测的可靠性和重复性，以供临床使用 可翻译的电动汽车测试。我们还将对化验设计和样品处理实施严格的质量控制， 收集经过良好注释的患者和对照样本，并进行统计支持的临床研究。这个 这项研究的技术和科学成果可能会对胶质瘤产生重大的翻译影响 通过建立一个强大和高度特异性的胶质瘤诊断和治疗监测的临床工具进行护理。

Malignant gliomas (MG) are the most aggressive malignancy of the central nervous system with the mean survival time of 15 months. Fast progression and high heterogeneity of MG mandates frequent imaging (MRI) to measure therapeutic responses. Imaging alone, however, is limited by false positives (“pseudoprogression”) and lacks molecular specificity. Tissue analyses could augment imaging results, but carries the risk of comorbidity and sampling errors. The goal of this project is to address such challenges and advance a minimally invasive, hence serially repeatable clinical assay specific to MG. We will exploit extracellular vesicles (EVs) as a new class of circulating cancer biomarker. Increasing number of studies evidence EVs' potential utility: these vesicles, are released from all cells, function as reliable cellular surrogates and reflect global tumor burden, overcoming limitations of tumor heterogeneity and sampling bias. We now seek to establish a translational EV assay for MG diagnosis and treatment monitoring, and compare its performance with gold standard imaging-based diagnostics. First, we will standardize EV assay protocols for clinical workflow. We will adopt our validated technologies: ExoLution, a clinical grade kit for EV isolation; Shahky, a high-throughput plasmonic instrument for EV protein analyses; and digital droplet PCR for highly sensitive EV mRNA detection. Leveraging the developmental and regulatory expertise of Exosome Diagnostics, we will make these platforms ready for translation into clinical diagnostic laboratories. Second, we will perform a targeted clinical study, critically assessing EVs' diagnostic and prognostic capacity for GBM. We will collect circulating EVs from GBM patients undergoing therapies and monitor serial changes of EV protein/mRNA profiles, particularly to detect the early sign of acquired resistance. We will compare results from EV assays and accompanying MRI, and build an integrative model for treatment monitoring. We formed a powerful multidisciplinary team to conduct these projects: Brain Tumor Center at Massachusetts General Hospital (MGH) which operates a vast biobank program of human clinical samples; Center for Systems Biology at MGH, a pioneer in developing new technologies for EV analyses; and Exosome Diagnostics, a de-facto industry leader in EV-based liquid biopsy with extensive experience in assay standardization. Collectively, the team has a track record of studying EVs' potential as a gliomas biomarker and has established optimal EV assays for molecular analyses. We will ensure assay reliability and reproducibility to deliver clinically translatable EV tests. We will also impose stringent quality controls on assay design and sample processing, accrue well-annotated patient and control samples, and perform statistically powered clinical studies. The technical and scientific outcomes of this research could have a significant translational impact in gliomas care by establishing a robust and highly specific clinical tool for gliomas diagnostics and treatment monitoring.