Digital Spatial Profiler Analysis Instrument

数字空间剖面分析仪

• 批准号: 10175562
• 负责人: PETER D. ADAMS
• 金额: $ 30.25万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2022-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175562
• 关键词: Aging; Cells; Collaborations; Data; Degenerative Disorder; Environment; Faculty; Formalin; Frozen Sections; Funding; Genes; Genomics; Heterogeneity; Histology; Homeostasis; Human; Immunofluorescence Immunologic; Immunohistochemistry; Institutes; Malignant Neoplasms; Medical; Messenger RNA; Morphology; Mus; Nerve Degeneration; Neurosciences; Normal tissue morphology; Pathologic; Physiological; Protein Analysis; Proteins; RNA; RNA analysis; RNA marker; Research; Research Personnel; Resolution; Suspensions; Technology; Time; Tissues; Tumor Tissue; Tumor-Derived; digital; flexibility; insight; instrument; interest; multiplex assay; nano-string; novel; protein biomarkers; single cell sequencing; time use; tissue stem cells; trait; transcriptome; tumor

PROJECT SUMMARY Sanford Burnham Prebys Medical Discovery Institute (SBP) hosts ~40 labs with wide-ranging interests, spanning normal tissue stem cells, tissue homeostasis and function, neuroscience, cancer, aging and degenerative diseases of aging, including neurodegeneration. A detailed analysis of cell – cell heterogeneity and spatial organization or disorganization across a multicellular tissue is key to understanding these physiological and pathological tissue states and functions. To this end, and to stay cutting-edge, SBP requests support for a Nanostring GeoMX Digital Spatial Profiler (DSP) to enable spatially resolved, digital quantitation of proteins or mRNAs in a highly multiplexed assay on tissue sections. The GeoMX DSP will bridge spatial analyses in SBP's Histology Core (by traditional immunohistochemistry or immunofluorescence) and single cell sequencing approaches in SBP's Genomics Core. The former gives spatial information on target distribution across a tissue or tumor, but is very low plex in that only a handful a targets can be analyzed at the same time. The latter is very high plex, detecting 1000s of expressed genes in a single cell, but gives no spatial information because it starts with a tissue/tumor-derived cell suspension. The GeoMX DSP yields spatial information across the tissue or tumor but is also very high plex, detecting dozens of proteins, 1000s or RNAs or even whole transcriptome simultaneously. Protein or RNA quantitation is performed within regions of interest (ROI), defined morphologically or by expression of up to 4 protein or RNA markers of interest. ROI are flexibly defined and can, for example, be a discrete region of the tissue or a subset of cells scattered throughout the tissue. This versatile instrument permits analysis of either formalin-fixed or fresh-frozen sections, human or mouse tissues, RNA or protein analyses and targeted or unbiased whole transcriptome analyses. These traits of spatial resolution, high plex analysis and versatility make GeoMX DSP a transformative technology for SBP's major users and others. This application is supported by 10 “major users” at SBP who will take 75% of the instrument's available use time (AUT). Like any novel transformative technology, GeoMX DSP will generate previously unattainable data and insights leading to novel hypotheses, new funding and collaborations. This will enhance the research environment at SBP and our faculty's interactions and collaborations with other researchers.

项目摘要 Sanford Burnham Prebys医学发现研究所（SBP）主持了40个具有广泛兴趣的实验室， 正常组织干细胞，组织稳态和功能，神经科学，癌症，衰老和退化性 衰老的疾病，包括神经变性。细胞 - 细胞异质性和空间的详细分析 多细胞组织的组织或混乱是了解这些生理和 病理组织状态和功能。为此，为了保持尖端，SBP请求支持 纳米弦GEOMX数字空间探测器（DSP）以实现空间解决，蛋白质的数字定量或 在组织切片上高度多重测定中的mRNA。 GEOMX DSP将在SBP中桥接空间分析 组织学核心（通过传统的免疫组织化学或免疫荧光）和单细胞测序 SBP基因组学核心的方法。前者提供有关跨组织目标分布的空间信息 或肿瘤，但很低的plex，因为只能同时分析少数靶标。后者非常 高PLEX，在单个单元格中检测1000秒的表达基因，但没有提供空间信息，因为它开始 具有组织/肿瘤衍生的细胞悬浮液。 GEOMX DSP在整个组织中产生空间信息或 肿瘤，但也很高的频谱，检测数十个蛋白质，1000秒或RNA甚至整个转录组 相似地。定义的蛋白质或RNA定量在感兴趣的区域（ROI）内进行 形态学或通过表达多达4种蛋白质或感兴趣的RNA标记。投资回报率灵活地定义，可以， 例如，是组织的离散区域或散布在整个组织中的细胞子集。这个多功能 仪器允许分析福尔马林固定或新鲜固定部分，人类或小鼠组织，RNA或 蛋白质分析，靶向或无偏的整体转录组分析。这些空间分辨率的特征，高 PLEX分析和多功能性使GEOMX DSP成为SBP主要用户和其他人的变革技术。 此应用程序由SBP的10个“主要用户”支持，他们将占用该仪器的75％ 时间（自动）。像任何新型的变革性技术一样，GEOMX DSP将生成以前无法实现的数据 以及导致新假设，新资金和合作的见解。这将增强研究 SBP的环境以及我们教师与其他研究人员的互动和合作。