Clinical translation of a NexGen platform for quantifying protein networks in human biospecimens

项目概要/摘要 由于技术限制，没有经验证的测定法用于定量人血清中的大多数人蛋白质。 临床生物样本，因此大多数人类蛋白质组在临床上是不可接近的。因为大多数现代 癌症治疗作用于蛋白质，我们无法量化蛋白质是一个障碍，以翻译的目标 治疗此外，蛋白质作为相互连接的“网络”，因此我们必须能够量化面板 癌症中的蛋白质，以评估决定治疗反应的途径和网络的活性。我们 项目旨在翻译（用于临床试验设置）用于定量肿瘤的多重蛋白质测定 抑制蛋白和细胞信号网络，响应DNA损伤-一个关键的网络， 目前的药物发现工作。该检测基于NextGen平台，用于定量蛋白质组使用 靶向多反应监测质谱（MRM-MS），补充现有的蛋白质测定 技术，并克服了许多技术限制，包括能够进行多重测定， 一个网络中的许多蛋白质。MRM已在临床前领域得到广泛验证，但尚未 已经被用于临床试验。我们的多学科团队汇集了学术合作伙伴， 专业知识（MRM-MS、分析化学、临床化学、病理学、统计学、生物工程、蛋白质 生物化学）与工业合作伙伴（阿斯利康，在药物开发方面具有丰富的经验， 临床试验这种基于MRM的检测面板成功适应临床试验环境将提供一条道路， 地图和标准操作协议（SOP），用于翻译额外的基于MRM的检测板（> 1，500 测定法已经在https：//assays.cancer.gov/公开获得）。

Project Summary/Abstract Due to technological limitations, there are no validated assays for quantifying the majority of human proteins in clinical biospecimens, and thus most of the human proteome is clinically inaccessible. Because most modern cancer therapies act on proteins, our inability to quantify proteins is an impediment to the translation of targeted therapies. Furthermore, proteins act as interconnected “networks,” and thus we must be able to quantify panels of proteins in cancers to assess the activity of pathways and networks that determine treatment responses. Our project aims to translate (for use in clinical trial settings) a multiplex protein assay for quantifying tumor suppressor proteins and cell signaling networks that response to DNA damage- a critical network targeted by current drug discovery efforts. The assay is based on a NextGen platform for quantifying protein panels uses targeted, multiple reaction monitoring mass spectrometry (MRM-MS), which complements existing protein assay technologies and overcomes many of the technological limitations, including enabling multiplexing of assays for many proteins in a single network. MRM has been extensively validated in the preclinical space but has not yet been adapted for clinical trials. Our multidisciplinary team brings together academic partners with diverse expertise (MRM-MS, analytical chemistry, clinical chemistry, pathology, statistics, bioengineering, protein biochemistry) with an industrial partner (AstraZeneca) that has extensive experience in drug development and clinical trials. Successful adaptation of this MRM-based assay panel to the clinical trial setting will provide a road map and standard operating protocols (SOPs) for translation of additional MRM-based assay panels (>1,500 assays are already publicly available at https://assays.cancer.gov/).