Rapid highly multi-plexed immuno-profiling of solid tumors by SpectralEdge imaging

项目摘要-摘要 免疫检查点抑制剂（ICI）的巨大影响引起了人们对研究的兴趣。 免疫-肿瘤细胞相互作用，以了解ICI敏感性和耐药性的机制， 对特定药物的反应，并开发新的治疗方法。多路组织成像是一种非常有前途的 肿瘤免疫分析方法：它可以评估许多细胞类型和状态， 肿瘤结构，并与病理学工作流程兼容。对多路复用存在一个关键的未满足的需求 该技术具有以下能力：（1）全载玻片成像;（2）快速单次数据收集;（3）子 细胞成像分辨率;（4）足够的免疫分析深度;（5）明场成像能力;（6）光学 和机械稳定性，用于精确的图像拼接;（7）核心和可定制抗体的可用性 用于轮廓分析的面板套件;以及（8）与已建立的图像数据标准和格式的兼容性。 这个RareCyte -哈佛医学院二期提案描述了SpectralEdgeTM，一个突破 成像技术，满足这些要求，执行高分辨率高复杂的全载玻片成像， 在不到1小时的单次扫描中使用荧光团缀合抗体标记20种抗原。的 专利的SpectralEdge方法涉及一种新型仪器和软件， TissuePlexTM antibody panel. SpectralEdge成像将加速发现预测和预后 生物标志物，使正在进行临床试验的免疫肿瘤药物的药效学研究成为可能， 最终提供临床上可行的诊断测试。 Aim 1将专注于设计和生产一个强大的，生产就绪的幻灯片扫描仪， 用于快速高分辨率20-plex SpectralEdge全载玻片成像的软件;它将基于RareCyte的 现有的Cyteplasmid II成像系统。生产研究专用仪器和试剂， ISO 13485标准将使后续部署在临床环境中成为可能。兼容当前和 新兴的数据格式和软件标准将能够与各种研究和临床 工作流。 AIM 2将测试和验证用于免疫表型和肿瘤细胞成像的模块化抗体组 在一系列肿瘤类型中。这将产生经验证的具有核心免疫谱集的组织学检测板 抗体和抗体“模块”。 目标3将在以下方面对充分表征的样本组测试SpectralEdge成像：（i）平移 生物标志物发现，（ii）行业赞助和制药商发起的免疫肿瘤学试验，（iii）小鼠 癌症模型，和（iv）目前正在用其他诊断方法进行免疫分析的临床标本 方法.临床样本研究将由委员会认证的HMS病理学家组成的咨询小组监督。

PROJECT SUMMARY – ABSTRACT The dramatic impact of immune checkpoint inhibitors (ICIs) has focused interest in investigating immune-tumor cell interactions to understand mechanisms of ICI sensitivity and resistance, to identify patients responsive to specific drugs, and to develop new therapies. Multiplexed tissue imaging is a highly promising approach to tumor immunoprofiling: it can assess many cell types and states within the context of preserved tumor architecture and is compatible with pathology workflows. A critical unmet need exists for multiplex technology with the following capabilities: (1) whole-slide imaging; (2) rapid single-pass data collection; (3) sub- cellular imaging resolution; (4) sufficient immunoprofiling depth; (5) bright-field imaging capability; (6) optical and mechanical stability for accurate image tile stitching; (7) availability of core and customizable antibody panel kits for profiling; and (8) compatibility with established image data standards and formats. This RareCyte – Harvard Medical School Phase II proposal describes SpectralEdgeTM, a breakthrough imaging technology that meets these requirements, performing high-resolution high-plex whole-slide imaging of 20 antigens labelled with fluorophore-conjugated antibodies in a single-pass scan of less than 1 hour. The patented SpectralEdge method involves a novel instrument and software as well as highly validated TissuePlexTM antibody panels. SpectralEdge imaging will accelerate discovery of predictive and prognostic biomarkers, enable pharmacodynamics study of immuno-oncology drugs undergoing clinical trials and ultimately provide clinically actionable diagnostic tests. Aim 1 will focus on engineering and production of a robust, production-ready slide scanner and software for rapid high-resolution 20-plex SpectralEdge whole-slide imaging; it will be based on RareCyte’s existing CyteFinder II imaging system. Production of a research-use-only instrument and reagents to ISO13485 standards will enable subsequent deployment in a clinical setting. Compatibility with current and emerging data format and software standards will enable integration with diverse research and clinical workstreams. Aim 2 will test and validate modular antibody panels for immunophenotyping and tumor cell imaging across a range of tumor types. This will yield validated TissuePlex panels having a core set of immunoprofiling antibodies and antibody “modules” for individual tumor types. Aim 3 will test SpectralEdge imaging on well-characterized sample cohorts in: (i) translational biomarker discovery, (ii) industry-sponsored and investigator-initiated immuno-oncology trials, (iii) mouse models of cancer, and (iv) clinical specimens currently undergoing immunoprofiling with other diagnostic methods. Clinical sample studies will be overseen by an advisory group of board-certified HMS pathologists.