Department of Cancer Immunology, Genentech, Inc., South San Francisco, CA, USA.
Department of Pathology, Genentech, Inc., South San Francisco, CA, USA.
J Pathol. 2019 Apr;247(5):650-661. doi: 10.1002/path.5223. Epub 2019 Feb 20.
Tumor cell heterogeneity and tumor cell-stromal interactions are being explored as determinants of disease progression and treatment resistance in solid tumor and hematological malignancies. As such, tools simultaneously capable of highly multiplexed profiling of tissues' protein and RNA content, as well as interrogation of rare or single cells, are required to precisely characterize constituent tumor cell populations, infiltrating lymphocytes and stromal elements. Access to spatial relationships will enable more precise characterization of tumors, support patient stratification and may help to identify novel drug targets. Multiple platforms are being developed to address these critical unmet needs. The NanoString digital spatial profiling (DSP) platform enables highly multiplexed, spatial assessment of protein and/or RNA targets in tissues by detecting oligonucleotide barcodes conjugated via a photocleavable linker to primary antibodies or nucleic acid probes. Although this platform enables high-dimensional spatial interrogation of tissue protein and RNA expression, a detailed understanding of its composition, function and chemistry is advisable to guide experimental design and data interpretation. The purpose of this review is to provide an independent, comprehensive description of the DSP technology, including an overview of NanoString's capture and antibody barcode conjugation chemistries, experimental workflow, data output and analysis methods. The DSP technology will be discussed in the context of other highly multiplexed immunohistochemistry methods, including imaging mass cytometry and multiplexed ion beam imaging, to inform potential users of the advantages and limitations of each. Additional issues such as preanalytical variability, sampling and specimen adequacy will be considered with respect to the platforms to inform potential experimental design. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
肿瘤细胞异质性和肿瘤细胞-基质相互作用正被探索为实体瘤和血液恶性肿瘤疾病进展和治疗耐药的决定因素。因此，需要能够同时高度多重分析组织蛋白和 RNA 含量，以及检测稀有或单个细胞的工具，以便精确描绘肿瘤细胞群体、浸润淋巴细胞和基质成分。获得空间关系将能够更精确地描述肿瘤，支持患者分层，并可能有助于识别新的药物靶点。正在开发多种平台来满足这些关键的未满足需求。NanoString 数字空间分析（DSP）平台通过检测与通过光裂解连接体连接的寡核苷酸条码来实现组织中蛋白和/或 RNA 靶标的高度多重化、空间评估，该连接体连接到初级抗体或核酸探针上。虽然该平台能够对组织蛋白和 RNA 表达进行高维空间检测，但为了指导实验设计和数据解释，建议对其组成、功能和化学性质有一个详细的了解。本综述的目的是提供对 DSP 技术的独立、全面描述，包括 NanoString 的捕获和抗体条码偶联化学、实验工作流程、数据输出和分析方法的概述。将在其他高度多重化免疫组织化学方法的背景下讨论 DSP 技术，包括成像质谱细胞术和多重离子束成像，以告知每种方法的优缺点。将考虑与平台相关的其他问题，如分析前变异性、采样和标本充足性，以告知潜在的实验设计。版权所有 © 2018 英国和爱尔兰病理学学会。由 John Wiley & Sons, Ltd. 出版
