Imaging and Liquid Biopsy for Glioma Diagnosis and Treatment Monitoring

恶性神经胶质瘤（MG）是中枢神经系统最具侵袭性的恶性肿瘤，平均 生存时间15个月。MG的快速进展和高度异质性要求频繁的成像（MRI） 来测量治疗反应。然而，单独的成像受到假阳性（“假进展”）的限制。 并且缺乏分子特异性。组织分析可以增强成像结果，但存在以下风险 协方差和抽样误差。该项目的目标是应对这些挑战， 微创，因此可连续重复的MG特异性临床试验。我们将利用细胞外 囊泡（EV）作为一类新的循环癌症生物标志物。越来越多的研究表明， 潜在效用：这些囊泡从所有细胞中释放出来，作为可靠的细胞替代物发挥作用， 全球肿瘤负荷，克服肿瘤异质性和采样偏倚的局限性。我们现在寻求 建立了用于MG诊断和治疗监测转化EV检测方法，并对其性能进行了比较 黄金标准的成像诊断。首先，我们将标准化EV检测方案， 工作流我们将采用我们经过验证的技术：ExoLution，一种用于EV分离的临床级试剂盒; Shahky，一种用于EV分离的临床级试剂盒。 用于EV蛋白分析的高通量等离子体仪器;以及用于高灵敏度EV的数字液滴PCR mRNA检测。利用外泌体诊断的开发和监管专业知识，我们将 使这些平台能够转化为临床诊断实验室。第二，我们将执行一个 有针对性的临床研究，严格评估EV对GBM的诊断和预后能力。我们将收集 来自接受治疗的GBM患者的循环EV，并监测EV蛋白/mRNA的系列变化 这是一个很好的方法，特别是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。我们建立了一个强大的 多学科团队进行这些项目：马萨诸塞州总医院脑肿瘤中心 (MGH)该中心运营着一个庞大的人类临床样本生物库项目; MGH是开发EV分析新技术的先驱; Exosome Diagnostics是事实上的 EV液体活检行业领导者，在检测标准化方面拥有丰富经验。统称 一个研究小组有研究EV作为神经胶质瘤生物标志物的潜力的记录，并建立了最佳EV 用于分子分析的测定。我们将确保检测的可靠性和重现性，以提供临床 可翻译的EV测试。我们还将对检测设计和样品处理实施严格的质量控制， 收集注释良好的患者和对照样本，并进行统计学上有效的临床研究。的 这项研究的技术和科学成果可能对神经胶质瘤产生重大的转化影响， 通过建立一个强大的和高度具体的临床工具，神经胶质瘤诊断和治疗监测护理。

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.