CODEX System for Automated and Highly Multiplexed Immunofluorescence

项目摘要 范德比尔特大学医学中心申请资金购买Akoya CODEX（通过indEXing进行CO检测） 包括CODEX射流仪器、CODEX处理计算机和Keyence显微镜的系统 使用过滤器，将其放置在免疫表型共享资源（IPSR）中。 组织微环境的空间分辨率，特别是免疫细胞浸润及其与 免疫和基质细胞，是癌症生物学、感染性疾病和自身免疫性疾病的基本方法， 免疫生物学调查，在临床前机制研究和翻译领域。法典体系 允许这种全面的、空间分辨的、高度多重的生物标志物分析（40多种抗原）， 福尔马林固定石蜡包埋或冷冻组织。这种高度多重的测定是通过缀合 将条形码化的寡核苷酸与抗体杂交，然后与染料标记的报道分子杂交，以获得高度特异性的寡核苷酸。 检测和有效的解链/再杂交/荧光检测循环。在2019年的范德比尔特糖尿病 研究和培训中心和生物分子多模态成像中心分别购买了一台CODEX 系统为自己的潜心研究。自购置以来，这些系统一直在满负荷运转， 吞吐量，以解决其他范德比尔特研究人员的研究。这种吞吐量的缺乏严重限制了 癌症研究人员和研究非糖尿病自身免疫性疾病的研究人员快速识别 新的生物标志物或组织中复杂的免疫状态。购买另一个CODEX系统将 大大缓解了这一研究瓶颈。 CODEX系统将支持NIH资助的8个主要用户和2个次要用户正在进行的研究， 范德比尔特大学和范德比尔特大学医学中心的多个部门。这些调查人员 迫切需要在细胞和亚细胞水平上进行高维原位成像分析，以研究宿主- 肿瘤相互作用、免疫细胞亚群以及对治疗的反应和抗性。除了这些 研究者项目，我们预计许多其他NIH资助的项目将需要CODEX系统，使 突破性的发现，特别是那些研究牛皮癣，类风湿性关节炎，肺部炎症（例如， SARS-CoV-2感染），以及其他可能进行有限活检的情况。 范德比尔特的CODEX系统的专业知识和支持是卓越的。IPSR拥有丰富的十年 研究宿主-肿瘤相互作用的杰出合作工作的历史，并有成功的实践经验， 在肿瘤组织上使用CODEX系统的经验。IPSR由三名既定教师共同领导 在执行高度复杂的免疫测定方面具有丰富的经验，这对成功的 实施CODEX系统。Young Kim博士（项目PI）和Jeff拉斯梅尔（主要用户），IPSR 科学总监和IPSR董事总经理Kim Dahlman博士，沿着经验丰富的IPSR研究团队 将为CODEX系统提供良好的指导、协助和管理。

Project Summary Vanderbilt University Medical Center requests funds to purchase an Akoya CODEX (CO-Detection by indEXing) System that includes the CODEX fluidics instrument, CODEX processing computer, and a Keyence microscope with filters, to be placed in the Immunophenotyping Shared Resource (IPSR). Spatial resolution of the tissue microenvironment, particularly immune cell infiltration and their relationships with immune and stromal cells, is an essential approach for cancer biology, infectious disease, and autoimmune immunobiology inquiries, in both preclinical mechanistic studies and translational domains. The CODEX System allows for this comprehensive, spatially resolved, highly multiplexed biomarker analysis (40+ antigens) on formalin-fixed paraffin-embedded or frozen tissue. This highly multiplexed assay is achieved by conjugating barcoded oligonucleotides to antibodies followed by hybridization with a dye-labeled reporter for highly specific detection and efficient melting/rehybridization/fluorescent detection cycles. In 2019 the Vanderbilt Diabetes Research and Training Center and the Biomolecular Multimodal Imaging Center each purchased a CODEX System for their dedicated studies. Since acquisition, the systems are operating at capacity and lack the throughput to address the research of other Vanderbilt investigators. This lack of throughput severely limits the capability of cancer researchers and researchers studying non-diabetic autoimmune diseases to rapidly identify novel biomarkers or complex immunological states in tissues. The purchase of another CODEX System will greatly alleviate this research bottleneck. The CODEX System will support the ongoing NIH-funded research of 8 Major Users and 2 Minor Users across multiple departments at Vanderbilt University and Vanderbilt University Medical Center. These investigators have a critical need for high-dimensional in situ imaging analysis at cellular and subcellular levels for studying host- tumor interactions, immune cell subsets, and response and resistance to therapies. In addition to these investigator projects, we anticipate that many other NIH-funded projects will require the CODEX System to make ground-breaking discoveries, particularly those studying psoriasis, rheumatoid arthritis, lung inflammation (e.g. SARS-CoV-2 infection), and others where limited biopsies are possible. The expertise and support for the CODEX System at Vanderbilt are exceptional. The IPSR has a rich ten-year history of outstanding collaborative work studying host-tumor interactions, and has successful hands-on experience using the CODEX System on tumor tissue. The IPSR is co-led by three established faculty members with a wealth of experience in performing highly complex immunological assays, which is critical to the successful implementation of the CODEX System. Drs. Young Kim (Project PI) and Jeff Rathmell (Major User), IPSR Scientific Directors, and Dr. Kim Dahlman, IPSR Managing Director, along with experienced IPSR research team will provide sound guidance, assistance, and management of the CODEX System.