Standardized Molecular Analyses of Glioma EVs

细胞外囊泡（EV）已成为组织活检的一种有前途的替代品， 非侵入性实时癌症监测大多数癌细胞释放大量EV进入循环， 携带反映母体肿瘤异质性的分子成分。该项目旨在 优化液体活检以诊断恶性神经胶质瘤。目前，这种肿瘤是通过 脑组织活检对患者有相当大的风险，不允许纵向随访， 临床护理目前的EV分离和表征方法产生不一致的结果和渲染数据 重现性具有挑战性，往往导致不可预测的结论。该项目的目标是i） 解决目前可用的不同EV隔离方法和平台之间的可变性，以及ii） 精确到“最佳”方法来验证胶质瘤诊断的候选生物标志物。我们卓越 调查小组汇集了恶性胶质瘤治疗，纳米工程领域， 囊泡研究，检测开发和液滴数字PCR技术，以优化必要的 用于开发能够走向临床环境的基于血液的测定的要素。通过 简单的血液检查，临床医生将能够诊断，分层和监测肿瘤，而不需要组织 活检我们与Exosome Diagnostics的战略合作伙伴关系，Exosome Diagnostics是EV癌症领域的行业领导者 诊断，为我们提供场地，允许我们的研究结果的翻译，加上获得临床级 试剂盒、平台和研究设计。神经外科学博士和系统生物学中心 马萨诸塞州总医院拥有多学科临床专业知识、创新技术和 互补资源，开展以下翻译项目：首先，基于我们以前的工具包 在对比工作中，我们选择了两个顶级的EV隔离套件和富集平台进行了一系列的测试 控制，参考标准，以确定最佳的EV隔离方法。 第二，我们将测试EV是否 基因标记可用作癌症检测以及追踪复发的生物标记。通过遵循 对器械设计和样本处理进行质量控制，获得注释良好的患者和对照样本， 并进行多中心测试，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诊断。通过文献数据挖掘，根据 他们可以区分神经胶质瘤和对照组。 最后，一个基因的签名与最高 灵敏度和特异性将在大量患者样品中得到验证。的技术和科学 这项研究的结果可能对神经胶质瘤产生重大的转化影响，建立一个强大的， 用于指导治疗决策和评估肿瘤复发的高度特异性测定。

Extracellular vesicles (EVs) have emerged as a promising surrogate for tissue biopsy, potentially enabling non-invasive, real-time cancer monitoring. Most cancer cells release large numbers of EVs into circulation that carry molecular constituents reflective of the heterogeneity of the parent tumor. This project is designed to optimize a liquid biopsy to diagnose malignant glioma tumors. Currently, such tumors are diagnosed through a brain tissue biopsy which involves considerable risk for patients and doesn’t allow for longitudinal follow up of clinical care. Current EV isolation and characterization methods yield inconsistent results and render data reproducibility challenging, often leading to unpredictable conclusions. The ​goals​ of this project are to i) address variability among the different EV isolation methods and platforms currently available, and to ii) pinpoint to the “best” method to validate candidate biomarkers for glioma diagnosis. Our exceptional investigative team brings together experts in malignant glioma treatment, the field of nano-engineering, vesicular research, assay development and droplet digital PCR technology to optimize the necessary elements for the development of a blood-based assay capable of moving towards clinical settings. Through a simple blood test, clinicians will be able to diagnose, stratify and monitor a tumor without the need for tissue biopsy. Our strategic partnership with ​Exosome Diagnostics​, an industry leader in EV-based cancer diagnostics, offers us venues allowing for the translation of our findings, coupled with access to clinical grade kits, platforms and study design. The D​ epartment of Neurosurgery​ and the ​Center for Systems Biology​ at the Massachusetts General Hospital comprise multidisciplinary clinical expertise, innovative technologies and complementary resources to carry out the following translational projects: ​First,​ based on our prior kit comparison work, we have picked two top EV isolation kits and enrichment platforms to test in a series of well controlled, reference standards to determine an optimal EV isolation method. ​Second,​ we will test whether EV gene signatures can be used as biomarkers for cancer detection as well as tracking recurrence. By following quality control on device design and sample processing, accruing well-annotated patient and control samples, and performing multi site testing, we will ​ensure assay reliability and reproducibility​ to deliver clinically translatable EV diagnostics. Fourteen genes were selected through literature data mining based on the putative evidence that they can distinguish gliomas from controls. ​Finally,​ a gene’s signature with the highest sensitivity and specificity will be validated in a large cohort of patient samples. The technical and scientific outcomes of this research could have a significant ​translational impact in gliomas​, establishing a robust, highly specific assay to guide treatment decision and assess tumor recurrence.