Advanced Cancer Classification via Single-Cell Electrophoretic Cytopathology

通过单细胞电泳细胞病理学进行高级癌症分类

基本信息

批准号：
9482979
负责人：
Amy Elizabeth Herr
金额：
$ 145.39万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9482979
关键词：
Advanced Malignant Neoplasm Antibodies Antineoplastic Agents Archives Benchmarking Binding Biological Biological Assay Biological Markers Biology Biometry Blood Vessels Breast Breast Cancer Treatment Cancer Diagnostics Cell Communication Cells Cellular Morphology Classification Classification Scheme Clinical Complex Cytometry Cytopathology Data Detection Development Diagnostic Drug resistance ERBB2 gene Epidermal Growth Factor Receptor Epitopes Exposure to Extracellular Domain Extracellular Matrix Fixatives Formalin Freezing Goals Heterogeneity Histology Human Hydrogels Image Immune Immunohistochemistry Individual Laboratories Length Malignant Neoplasms Mammary Neoplasms Measurement Measures Mesenchymal Methods Microfluidics Molecular Morphology Neoplasm Metastasis Oncoproteins Paraffin Embedding Pathology Patients Pharmaceutical Preparations Physical shape Post-Translational Protein Processing Precision therapeutics Progression-Free Survivals Protein Isoforms Protein Tyrosine Kinase Proteins Resistance Resolution Retrospective Studies Role Selection for Treatments Signal Transduction Signaling Protein Slice Statistical Methods Stromal Cells Taxonomy Technology Temperature Tissue Embedding Tissue Sample Tissues Trastuzumab Ultraviolet Rays Variant Western Blotting Woman anticancer research base biobank cancer classification cancer subtypes cancer therapy clinical Diagnosis commercialization design extracellular humanized monoclonal antibodies imprint improved innovation insight link protein malignant breast neoplasm mortality neoplastic cell next generation novel novel therapeutics personalized medicine prognostic protein biomarkers protein profiling resistance mechanism screening targeted cancer therapy therapy development tissue processing tool treatment strategy tumor tumor heterogeneity tumor microenvironment tumor progression

项目摘要

ABSTRACT Recent efforts to identify and circumvent mechanisms of drug resistance point to the role of HER2 truncated isoforms, which lack the extracellular domain of the full-length protein targeted by trastuzumab. Understanding the biology of HER2 isoforms is crucial to developing new drugs personalized for individuals resistant to trastuzumab therapy. However, detection of these isoforms in paraffin-embedded tissue sections (FFPE) is impossible by conventional methods (Immunohistochemistry, IHC) due to the lack of isoform-specific antibodies and the loss of epitope integrity during tissue processing. To overcome these barriers, the goal of this project is to develop a robust tool that will both allow retrospective studies on HER2 isoforms and enable isoform screening on a patient-to-patient basis to tailor breast cancer treatment. Our efforts fill a major gap in cancer research: the limited ability to scrutinize tissue sections for protein targets that either lack highly specific antibody probes (analytical) or lose epitope integrity during tissue processing (pre- analytical). We introduce a whole-tissue western blot (wtWestern) that creates a novel 3D protein ‘imprint’ of the tissue section for stable storage and downstream antibody probing. The unique dual-functionality wtWestern electrophoretically resolves proteins (extracted from a 2D tissue section) along the depth (z-axis) of a supporting photoactive hydrogel layer. Exposure to UV light covalently links protein to the hydrogel. We benchmark the fixative-free imprint for biobanking suitability, apply the wtWestern to measurement of proteins (including isoforms) in archived frozen or FFPE tissue sections, and maximize tissue utility by enabling detection of 20+ proteins for cancer sub-type classification. This innovative wtWestern is expected to maximize the quality and utility of biospecimens for downstream analyses – critically important to retrospective studies that form the basis for both cancer diagnostics/prognostics and personalized medicine.

抽象的识别和规避耐药性机制的最新努力表明HER2的作用已截断同工型缺乏由曲妥珠单抗靶向的全长蛋白的细胞外结构域。理解 HER2同工型的生物学对于开发针对耐心的个性化的新药物至关重要曲妥珠单抗治疗。但是，在石蜡包裹的组织切片（FFPE）中检测这些同工型是由于缺乏同工型特异性抗体，不可能通过常规方法（免疫组织化学，IHC）以及组织加工过程中表位完整性的丧失。为了克服这些障碍，该项目的目标是开发一种可靠的工具，该工具既可以允许对HER2同工型进行回顾性研究，又可以启用在患者到患者的基础上筛查同工型以量身定制乳腺癌治疗。我们的努力填补了癌症研究的主要空白：仔细检查蛋白质靶标的组织切片的能力有限在组织加工过程中缺乏高度特异的抗体问题（分析）或失去表位完整性（前分析）。我们介绍了一个全组织Western印迹（WTWestern），该印迹创建了一种新颖的3D蛋白“烙印” 稳定储存和下游抗体探测的组织截面。独特的双功能 wtwestern沿着深度（z轴）的蛋白质（从2D组织截面提取）（Z轴）（Z轴）支持光活性水凝胶层。暴露于紫外线将蛋白质与水凝胶联系起来。我们基准为生物库的固定烙印，以适用于蛋白质的测量（包括同工型）在存档的冷冻或FFPE组织切片中，以及通过启用检测的最大组织实用程序用于癌症亚型分类的20+蛋白。这种创新的WTWestern有望最大化质量生物测量对下游分析的实用性 - 对于形成形成的回顾性研究至关重要癌症诊断/预后和个性化医学的基础。