ShEEP Request for Imaging Mass Cytometry

ShEEP 成像质量细胞计数请求

• 批准号: 9905867
• 负责人: AMANDA C. LARUE
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9905867
• 关键词: Ablation; Address; Alzheimer' s Disease; Animal Model; Animals; Antibodies; Architecture; Behavioral; Bioinformatics; Biomedical Research; Cardiovascular Diseases; Cell Separation; Cells; Cellular Morphology; Clinical; Clinical Trials; Complement; Complex; Consultations; Core Facility; Cytometry; Data Analyses; Detection; Development; Disease; Disease Progression; Evaluation; Event; Extracellular Matrix; Feedback; Fees; Flow Cytometry; Genomics; Goals; Health; Histology; Human; Image; Immunology; In Situ; Injury; Isotopes; Label; Laboratories; Lasers; Logistics; Maintenance; Medical; Medical center; Mental disorders; Metals; Methodology; Methods; Molecular; Orthopedics; Pathologic; Pathologic Processes; Pathology; Patients; Physiological; Physiological Processes; Population Analysis; Positioning Attribute; Precision therapeutics; Proteins; Protocols documentation; Research; Research Personnel; Resolution; Resource Sharing; Sampling; Schedule; Services; Sheep; South Carolina; Spinal cord injury; Stem cells; Strategic Planning; Stroke; Structure; System; Technology; Time; Tissue Sample; Tissues; Training; Translations; Traumatic Brain Injury; Universities; Veterans; base; cell type; conflict resolution; design; dimensional analysis; experience; health care quality; high dimensionality; human disease; imaging capabilities; imaging modality; improved; innovative technologies; instrumentation; interest; next generation; novel; novel marker; novel therapeutics; oncology; operation; preservation; respiratory; tool; transcriptomics; translational medicine; treatment response

The goal of this proposal is to install and integrate the Fluidigm Hyperion Imaging Mass Cytometer (IMC) system in the Cellular and Molecular Evaluation Core (CMEC) Facility within Research Service at the Ralph H. Johnson (RHJ) VA Medical Center. The overarching objective of this core facility is to provide tools and services, including immunohistochemical, flow cytometry, and cell sorting applications, necessary to support and advance research that directly impacts the health of our Veterans. Conventional immunohistochemical (IHC) approaches have been historically limited to the simultaneous detection of only a few markers (~8). Traditional flow cytometry, while allowing for multiparameter analysis, does not allow for spatial analysis of cellular or subcellular marker expression within a tissue. In contrast, time of flight mass cytometry (CyTOF) allows for the simultaneous detection of 40+ markers on one single cell. By combining the existing high dimensional Helios mass cytometer CyTOF platform with the Hyperion laser ablation system, we are able to bring this powerful detection method, termed imaging mass cytometry (IMC), into tissues. The central hypothesis addressed by this core is that the high dimensional analysis Hyperion IMC system will allow VA researchers to use small fixed tissue samples to examine complex subcellular, cellular, and cell-matrix interactions in the intact tissue microenvironment and provide a technology bridge to existing transcriptomic and imaging methods to understand pathologies impacting our Veterans. Our investigators have identified four primary aims that will be accomplished through this groundbreaking technology that provides high throughput, high-dimensional quantitative analyses in situ of complex cellular samples: 1) Identify novel cell types and their cellular and sub-cellular proteins that contribute to the pathology of disease; 2) Uncover new intracellular, cell- cell, and cell-ECM interactions important for disease progression; 3) Examine rare events (including stem cells) in physiological and pathological conditions; and 4) Discover novel biomarkers of disease and/or clinical response to treatment, enhancing the development of targeted, personalized, precision therapies. The state-of-the-art technology afforded by the Hyperion IMC System is broadly applicable across the diverse animal- and human-based research efforts of our local VAMC investigators including studies in traumatic brain injury, stroke, spinal cord injury, respiratory illness, cardiovascular disease, immunology, oncology, orthopedic injury, Alzheimer’s disease, and mental health disorders. This technology does not currently exist at RHJ VAMC, the Affiliate (Medical University of South Carolina, MUSC), or in the state. Based on the number of interested investigators and diverse disease-related applications, we believe that this upgrade will meet demands of investigators for cutting edge technologies to advance translational medicine and allow the VA CMEC to provide the next generation of mass cytometry capabilities. This expansion will position VA as a leader in integration of mass cytometry into clinical trials, personalized disease treatments, and development of novel therapies. Incorporating the Hyperion Imaging Mass Cytometer System into the established CMEC will greatly strengthen the research capabilities of our station (and those in the VISN), facilitating basic scientific discovery of mechanisms underlying physiological and pathological processes and translation of these findings to support the rapid implementation of cutting-edge personalized medical treatment to improve quality healthcare for our Nation’s Veterans.

该提案的目标是安装和集成 Fluidigm Hyperion 成像质谱流式细胞仪 (IMC) 系统 拉尔夫·约翰逊 (Ralph H. Johnson) 研究服务部细胞和分子评估核心 (CMEC) 设施 (RHJ) VA 医疗中心 该核心设施的首要目标是提供工具和服务，包括 支持和推进研究所必需的免疫组织化学、流式细胞术和细胞分选应用 直接影响我们退伍军人的健康。 传统的流式细胞术历来仅限于同时检测少数标记物 (~8)。 虽然允许多参数分析，但不允许对细胞或亚细胞标记进行空间分析 相反，飞行时间质谱细胞术 (CyTOF) 允许同时进行。 结合现有的高维 Helios 质谱流式细胞仪，可检测单个细胞上的 40 多个标记。 CyTOF平台配合Hyperion激光烧蚀系统，我们能够带来这种强大的检测方法， 称为成像质量细胞术（IMC），进入组织。 该核心提出的中心假设是，高维分析 Hyperion IMC 系统将 允许 VA 研究人员使用小型固定组织样本来检查复杂的亚细胞、细胞和细胞基质 完整组织微环境中的相互作用，并为现有的转录组学和 我们的研究人员已经确定了四种影响退伍军人的病理学成像方法。 通过这种提供高吞吐量的突破性技术将实现的主要目标， 复杂细胞样品的原位高维定量分析：1) 识别新的细胞类型及其 有助于疾病病理学的细胞和亚细胞蛋白；2) 发现新的细胞内、细胞- 细胞和细胞-ECM 相互作用对疾病进展很重要；3) 检查罕见事件（包括干细胞） 生理和病理条件下；4) 发现疾病和/或临床的新生物标志物 对治疗的反应，促进有针对性的、个性化的、精准疗法的开发。 Hyperion IMC 系统提供的最先进技术广泛适用于各种领域 我们当地 VAMC 研究人员基于动物和人类的研究工作，包括脑外伤研究 损伤、中风、脊髓损伤、呼吸系统疾病、心血管疾病、免疫学、肿瘤学、骨科 RHJ VAMC 目前不存在这项技术， 附属机构（南卡罗来纳医科大学，MUSC），或州内的基于感兴趣的数量。 研究人员和各种疾病相关的应用程序，我们相信这次升级将满足 研究人员寻求尖端技术来推进转化医学，并允许 VA CMEC 提供 下一代质谱流式细胞仪功能的扩展将使 VA 成为集成的领导者。 质谱流式细胞术应用于临床试验、个性化疾病治疗和新疗法的开发。 将Hyperion成像质谱仪系统纳入已建成的CMEC将大大加强 我们站（以及 VISN 中的研究能力）的研究能力，促进了基础科学发现 生理和病理过程的潜在机制以及这些发现的转化以支持 快速实施尖端的个性化医疗，以提高我们的医疗质量 国家退伍军人。