ShEEP Request for Imaging Mass Cytometry

该计划的目标是安装和集成Fluidigm Hyperion成像质量细胞仪(IMC)系统 在拉尔夫·H·约翰逊研究所的细胞和分子评估核心(CMEC)设施中 (RHJ)退伍军人医疗中心这一核心设施的总体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类应用 这直接影响到我们退伍军人的健康。传统的免疫组织化学(IHC)方法 历史上仅限于同时检测几个标记(~8个)。传统的流式细胞术， 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 在组织中的表达。相比之下，飞行时间质量细胞术(CyTOF)允许同时 在单个细胞上检测40+个标记。通过结合现有的高维Helios质量细胞仪 CyTOF平台配合Hyperion激光消融系统，我们能够带来这种强大的检测方法， 被称为成像质量细胞术(IMC)，进入组织。 这个核心提出的中心假设是，高维分析Hyperion IMC系统将 允许退伍军人事务部研究人员使用小的固定组织样本来检查复杂的亚细胞、细胞和细胞基质 在完整的组织微环境中的相互作用，并提供了一座连接到现有转录和 了解影响退伍军人的病理的成像方法。我们的调查人员已经确认了四个 将通过这一突破性技术实现的主要目标是提供高吞吐量、 复杂细胞样本的高维原位定量分析：1)识别新的细胞类型和它们的 有助于疾病病理的细胞和亚细胞蛋白质；2)发现新的细胞内、细胞- 细胞、细胞与细胞外基质的相互作用对疾病进展很重要；3)检查罕见事件(包括干细胞) 在生理和病理条件下；以及4)发现疾病和/或临床的新生物标志物 应对治疗，促进有针对性、个性化、精准治疗的发展。 Hyperion IMC系统提供的最先进技术广泛适用于各种 我们当地VAMC研究人员的动物和人类研究努力，包括对创伤性脑的研究 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 伤害、阿尔茨海默氏症和精神健康障碍。这项技术目前在RHJ VAMC中不存在， 附属机构(南卡罗来纳医科大学，MUSC)，或在该州。根据感兴趣的人数 调查人员和多样化的疾病相关应用，我们相信这次升级将满足 尖端技术的调查人员推动转化医学，并允许退伍军人事务部提供 下一代的质量细胞仪能力。这一扩展将使退伍军人管理局成为整合 将质量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成像质量细胞仪系统纳入已建立的CMEC将大大加强 我们空间站(和VISN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翻译支持 快速实施尖端个性化医疗，提高我国医疗质量 国家的退伍军人。

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.