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: 10019483
• 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 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019483
• 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; improved; innovation; instrumentation; light microscopy; malignant breast neoplasm; mass spectrometric imaging; 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 在过去的三十年里，质谱学在蛋白质组学领域发挥了主导作用。一种组合 基于MS的创新技术为蛋白质组发现提供了强大的工具，包括 确定复杂样本中的蛋白质生物标记物，量化蛋白质表达的变化，并表征蛋白质- 蛋白质的相互作用。MS的第二个重要进展是引入了质谱学成像 (MSI)，它将MS扩展到空间维度，允许绘制生物分子的分布，包括 蛋白质、核酸、代谢物，甚至复杂组织中的小药物化合物。这一阶段的目标是 项目是进一步开发MSI执行高度多路复用成像的能力，即使是在亚细胞上 生物样品中的目标生物分子的纳米级。这样的能力将为系统提供一个主要工具 生物学家和癌症研究人员，他们需要详细了解复合体中关键分子的分布 在细胞、亚细胞和分子水平上的组织。它还将为病理学家提供一种强大的新工具 分析肿瘤组织标本，以最终获得改善的治疗和患者结果。在.期间 第一阶段我们已经成功地演示了：i)通过MSI同时成像的能力，可能有数百个 福尔马林固定石蜡包埋(FFPE)小鼠脑切片靶向生物标志物的研究 专有的改进型光裂解质谱标签(IPC-MTS)，它被结合到抗体或 寡核苷酸探针。相比之下，传统的基于光学显微镜的免疫组织化学(IHC)和 荧光原位杂交(FISH)方法只能对少数目标生物分子成像；以及ii)设计 新的IPC-MT-探针完全兼容新的膨胀显微镜(EXM)方法，以获得 纳米级亚细胞MSI分辨率。在第二阶段，我们将通过应用IPC-MT-Probe在此基础上再接再厉 对存档的乳腺癌FFPE样本进行分析的技术，以最终实现癌症的改善 常规临床实践的组织病理学。作为模型系统，10个IPC-MT-抗体探针和10个IPC-MT-抗体探针 将开发针对特定乳腺癌肿瘤抗原和miRNA生物标记物的寡核苷酸探针 并使用AmberGen专有的珠阵列质谱初步测试了潜在的交叉反应 技术(珠-AMS™)。然后，乳腺癌组织切片将使用这些探针和MSI进行分析 与传统的基于荧光的IHC和FISH方法所获得的结果进行比较。为了获得 更高的空间分辨率，IPC-MT-与扩展的丙烯酸酯凝胶交联或传递到扩展的丙烯酸酯凝胶的探针 将使用包含扩展的FFPE切片。膨化凝胶的MSI将使用专门的低- 温度、红外激光为主的MALDI-MSI仪器。这项工作将由我们继续 与MS和ExM领域的领先专家合作，包括Cathy Costello博士(BU，William Fairfield 沃伦著名教授，波士顿大学生物医学质谱学中心主任)，Ed Boyden(麻省理工学院，Y。 Eva Tan神经技术教授)和Jason Amsden(杜克大学助理研究教授)。

项目成果

Highly Multiplexed Immunohistochemical MALDI-MS Imaging of Biomarkers in Tissues.

• DOI: 10.1021/jasms.0c00473
• 发表时间: 2021-04-07
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Yagnik G;Liu Z;Rothschild KJ;Lim MJ
• 通讯作者: Lim MJ

Evaluation of antibody-based single cell type imaging techniques coupled to multiplexed imaging of N-glycans and collagen peptides by matrix-assisted laser desorption/ionization mass spectrometry imaging.

• DOI: 10.1007/s00216-023-04983-2
• 发表时间: 2023-11
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Dunne, Jaclyn;Griner, Jake;Romeo, Martin;Macdonald, Jade;Krieg, Carsten;Lim, Mark;Yagnik, Gargey;Rothschild, Kenneth J.;Drake, Richard R.;Mehta, Anand S.;Angel, Peggi M.
• 通讯作者: Angel, Peggi M.