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 过去三十年来，质谱（MS）在蛋白质组学领域发挥了主导作用。 基于 MS 的创新技术为蛋白质组学发现提供了强大的工具，包括能够 识别复杂样品中的蛋白质生物标志物，量化蛋白质表达的变化并表征蛋白质 蛋白质相互作用的第二个重要进展是质谱成像的引入。 (MSI) 将 MS 扩展到空间维度，可以绘制生物分子的分布图，包括 复杂组织中的蛋白质、核酸、代谢物甚至小药物化合物是第二阶段的目标。 该项目旨在进一步开发 MSI 执行高度多重成像的能力，甚至是在亚细胞上 纳米级的生物样本中的目标生物分子的这种能力将为系统提供主要工具。 生物学家和癌症研究人员，他们需要详细了解复杂的关键分子的分布 它也将为病理学家提供一个强大的新工具。 分析肿瘤组织标本，以便最终获得改善的治疗和患者结果。 第一阶段我们已经成功证明了：i) MSI 能够同时对数百个图像进行成像 使用来自小鼠大脑的福尔马林固定石蜡包埋 (FFPE) 薄片的靶向生物标志物 专有的改进型光裂解质量标签 (iPC-MT)，可整合到抗体中或 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进行成像；ii) 设计； 新的 iPC-MT 探针与扩展显微镜 (ExM) 新方法完全兼容，以获得 在第二阶段，我们将通过应用 iPC-MT 探针来实现纳米级亚细胞 MSI 分辨率。 技术用于分析存档的乳腺癌 FFPE 标本，以最终实现癌症的改善 作为常规临床实践的组织病理学模型系统，10 个 iPC-MT- 抗体探针和 10 个 iPC-MT- 将开发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 并使用 AmberGen 专有的珠阵列质谱法初步测试了潜在的交叉反应性 然后使用这些探针通过 MSI 技术 (Bead-AMS™) 分析乳腺癌组织切片。 与基于传统荧光的 IHC 和 FISH 方法获得的结果进行比较，以获得 提高空间分辨率，与膨胀丙烯酸酯凝胶交联或传递至膨胀丙烯酸酯凝胶的 iPC-MT 探针 将使用含有 FFPE 切片的扩展凝胶，并使用专门的低密度材料来实现。 温度、基于红外激光的 MALDI-MSI 仪器将通过我们的持续推动来促进。 与MS和ExM领域的领先专家合作，包括Cathy Costello博士（BU，William Fairfield） 沃伦杰出教授、波士顿大学生物医学质谱中心主任）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.

组织中生物标志物的高度多重免疫组织化学 MALDI-MS 成像。

• 发表时间: 2021-04-07
• 影响因子: 3.2
• 作者: Yagnik, Gargey;Liu, Ziying;Rothschild, Kenneth J;Lim, Mark J
• 通讯作者: Lim, Mark J

Real-time glioblastoma tumor microenvironment assessment by SpiderMass for improved patient management.

SpiderMass 实时评估胶质母细胞瘤肿瘤微环境，以改善患者管理。

• 发表时间: 2024-04-16
• 作者: Zirem, Yanis;Ledoux, Léa;Roussel, Lucas;Maurage, Claude Alain;Tirilly, Pierre;Le Rhun, Émilie;Meresse, Bertrand;Yagnik, Gargey;Lim, Mark J;Rothschild, Kenneth J;Duhamel, Marie;Salzet, Michel;Fournier, Isabelle
• 通讯作者: Fournier, Isabelle

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.

通过基质辅助激光解吸/电离质谱成像评估基于抗体的单细胞类型成像技术与 N-聚糖和胶原蛋白肽多重成像的结合。

• 发表时间: 2023-11
• 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.