Highly Multiplexed Nanoscale Mass Spectrometric Imaging of Cancer Tissues

癌症组织的高度多重纳米级质谱成像

• 批准号: 9908822
• 负责人: Mark Lim
• 金额: $ 99.97万
• 依托单位: AMBERGEN, INC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908822
• 关键词: Acrylates; Amines; Antibodies; Archives; Area; Binding; Biological Assay; Biological Markers; Biological Models; Brain; Collaborations; Complex; Cross Reactions; Data; Detection; Dimensions; Drug Compounding; Evaluation; Fluorescence; Fluorescent in Situ Hybridization; Formalin; Gel; Goals; Histopathology; Image; Immunohistochemistry; Knowledge; Label; Lasers; Letters; Malignant Neoplasms; Mass Spectrum Analysis; Methods; MicroRNAs; Microscopy; Microtomy; Molecular; Mus; Nucleic Acids; Oligonucleotide Probes; Oligonucleotides; Paraffin Embedding; Pathologic; Pathologist; Pathology; Patient-Focused Outcomes; Patients; Peptides; Phase; Physics; Play; Polymers; Proteins; Proteomics; Reagent; Research Assistant; Research Personnel; Resolution; Role; Sampling; Slice; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Statistical Methods; System; Techniques; Technology; Testing; Tissue Embedding; Tissue imaging; Tissues; Tumor Antigens; Tumor Tissue; Universities; Validation; Work; base; biological systems; cancer imaging; clinical practice; cold temperature; cross reactivity; crosslink; cryogenics; design; distinguished professor; imaging potential; improved; innovation; instrumentation; light microscopy; malignant breast neoplasm; microRNA biomarkers; molecular subtypes; multiplexed imaging; nanoscale; neurotechnology; novel; optical imaging; optimal treatments; professor; protein biomarkers; protein expression; protein protein interaction; targeted biomarker; therapy outcome; tool

Summary/Abstract Mass spectrometry (MS) has played a leading role in the past three decades in the field of proteomics. A combination of innovative MS-based techniques has provided powerful tools for proteomic discovery including the ability to identify protein biomarkers in a complex sample, quantify changes in protein expression and characterize protein- protein interactions. A second important advance in MS has been the introduction of mass spectrometric imaging (MSI) which extends MS to the spatial dimension, allowing mapping of the distribution of biomolecules including proteins, nucleic acids, metabolites and even small drug compounds in complex tissues. The goal of this Phase II project is to further develop the ability of MSI to perform highly multiplexed imaging, even on the subcellular nanoscale, of targeted biomolecules in biospecimens. Such a capability would provide a major tool for systems biologists and cancer researchers, who require a detailed knowledge of the distribution of key molecules in complex tissues at the cellular, subcellular and molecular levels. It would also provide pathologists with a powerful new tool to analyze tumor tissue specimens in order to ultimately obtain improved therapy and patient outcomes. During Phase I we have successfully demonstrated: i) the ability to simultaneously image by MSI potentially hundreds of targeted biomarkers from formalin fixed paraffin embedded (FFPE) thin sections from mouse brain using proprietary improved photocleavable mass-tags (iPC-MTs) which are incorporated into antibodies or oligonucleotide probes. In contrast, conventional light microscopy-based immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) methods can image only a few targeted biomolecules; and ii) designed new iPC-MT-probes which are fully compatible with the new method of expansion microscopy (ExM) to obtain nanoscale subcellular MSI resolution. During Phase II, we will build on this progress by applying iPC-MT-probe technology to the analysis of archived breast cancer FFPE specimens in order to ultimately achieve improved cancer histopathology for routine clinical practice. As a model system, 10 iPC-MT-antibody probes and 10 iPC-MT- oligonucleotide probes targeted at specific breast cancer tumor antigens and miRNA biomarkers will be developed and initially tested for potential cross-reactivity using AmberGen's proprietary Bead-Array Mass Spectrometry technology (Bead-AMS™). Breast cancer tissue slices will then be analyzed by MSI using these probes and compared to results obtained from conventional fluorescence based IHC and FISH methods. In order to obtain increased spatial resolution, iPC-MT-probes that cross-link with or are delivered to expanded acrylate gels containing expanded FFPE slices will be used. MSI of the expanded gels will be achieved using specialized low- temperature, infrared laser-based MALDI-MSI instrumentation. This work will be facilitated by our continued collaboration with leading experts in the MS and ExM fields including Drs. Cathy Costello (BU, William Fairfield Warren Distinguished Professor, Director of BU Center for Biomedical Mass Spectrometry), Ed Boyden (MIT, Y. Eva Tan Professor of Neurotechnology) and Jason Amsden (Duke University, Assistant Research Professor).

摘要/摘要 质谱法（MS）在过去的蛋白质组学领域中发挥了领导作用。 创新的基于MS的技术已经为蛋白质组学提供了强大的工具，包括 鉴定复杂样品中蛋白质生物标志物，量化蛋白质表达的变化并表征蛋白质 - 蛋白质相互作用。 （MSI）将MS扩展到空间维米座，允许映射生物分子的分布 蛋白质，核酸，代谢产物和综合组织中的小型药物化合物。 项目将进一步发展MSI执行高度多路复用成像的能力，即使在亚细胞上也是如此 纳米级的生物分子中的生物分子。 生物学家和癌症研究人员，他们需要对复杂分子的分布进行详细了解 细胞，亚细胞和分子水平的组织。 分析肿瘤组织标本，以最终获得改善的治疗和患者的结局。 第一阶段我们已经成功证明了：i）MSI同时形象的能力可能有数百个 使用福尔马林固定石蜡嵌入（FFPE）的靶向生物标志物使用小鼠脑的薄切片 专有改进的光电标签（IPC-MTS）掺入抗体或 寡核苷酸探针，基于光学显微镜的免疫组织化学（IHC）和 荧光原位杂交（FISH）方法只能成像少数目标生物分子； 与新的扩展显微镜（EXM）完全兼容的新IPC-MT探针可获得 纳米级亚细胞MSI分辨率。 用于分析存档乳腺癌FFPE标本的技术，以最终实现癌症 常规临床实践的组织病理学。 针对特定乳腺癌肿瘤抗原和miRNA生物标志物的寡核苷酸探针将是开发 并最初使用Ambergen的专有珠阵列质谱法测试了潜在的交叉反应性 技术（Bead-AMS™）。 与传统的基于荧光的IHC和FISH方法获得的结果相比。 增加空间分辨率，与或交付到扩展的丙烯酸酯凝胶的IPC-MT探针 将使用包含扩展的FFPE切片。 温度，红外激光的MALDI-MSI仪器将通过我们的续 与MS和EXM领域的主要专家合作，包括Drs（BU，William Fairfield） 沃伦（Warren）杰出教授，埃德·博伊登（Ed Boyden）Bu Bu Bu Bu Bu Bu Centrals的Bu Bu Bu Bu Centrals（Mit，Y。） EVA TAN神经技术教授）和Jason Amsden（杜克大学，助理研究教授）。