Standardized Molecular Analyses of Glioma EVs

细胞外小泡(EVS)已成为一种很有前途的组织活检替代物，有可能使 非侵入性的、实时的癌症监测。大多数癌细胞释放大量的EV进入血液循环 携带反映亲本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胶质瘤肿瘤。目前，这种肿瘤的诊断是通过一种 脑组织活检对患者有相当大的风险，不允许进行纵向随访 临床护理。当前的EV分离和表征方法产生不一致的结果和呈现数据 重复性具有挑战性，经常导致不可预测的结论。这个项目的​Goals​是to i) 解决目前可用的不同EV隔离方法和平台之间的差异，以及 精确到“最佳”方法，以验证胶质瘤诊断的候选生物标记物。我们的非凡之处 研究小组汇集了恶性胶质瘤治疗、纳米工程领域的专家， 水泡研究、化验开发和水滴数字聚合酶链式反应技术的优化 开发一种能够应用于临床的基于血液的检测方法的要素。通过一个 简单的血液测试，临床医生将能够诊断、分层和监测肿瘤，而不需要组织 活组织检查。我们与基于EV的癌症行业领先者​Exosome Diagnostics​建立了战略合作伙伴关系 诊断学，为我们提供了翻译我们的研究结果的场所，以及获得临床分级的机会 工具包、平台和书房设计。神经外科D​分部和​系统生物学中心​ 马萨诸塞州综合医院包括多学科的临床专业知识、创新技术和 补充资源以执行以下翻译项目：​First，基于我们以前的工具包的​ 对比工作中，我们选择了两个顶级的电动汽车隔离套件和浓缩平台在一系列油井中进行测试 受控的参考标准，以确定最佳的EV隔离方法。​Second，​我们将测试电动汽车 基因签名可用作癌症检测和跟踪复发的生物标记物。通过以下方式 对仪器设计和样品处理的质量控制，收集经过良好注释的患者和对照样品， 并进行多点测试，我们将​确保分析的可靠性和重复性​提供给临床 可翻译的电动汽车诊断。在此基础上，通过文献数据挖掘筛选出14个基因 他们可以区分胶质瘤和对照组的推定证据。​最后，​一个基因的签名具有最高的 敏感度和特异度将在大量患者样本中得到验证。技术和科学 这项研究的结果可能对​在胶质瘤​中的翻译产生重大影响，建立一个强大的， 用于指导治疗决策和评估肿瘤复发的高特异性检测。

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.