Transparent Tumor Tomography (T3): Multi-Parameter 3D Imaging for Tumor Immunotherapy

项目摘要 该提案旨在将成像技术应用于三维(3D)可视化和地图绘制 肿瘤和微环境，包括癌细胞、间质细胞、免疫细胞和血管细胞。这个 能够确定不同细胞群的位置、密度和功能方向 肿瘤不仅将是癌症生物学的有力工具，而且将是改善诊断和推进癌症治疗的有力工具。 开发有效的治疗方法。虽然免疫组织化学(IHC)/免疫荧光(IF)染色 长期以来一直被用于癌症诊断，传统的IHC/IF方法是基于本地化的 单个薄片中的抗原。考虑到视场中的大多数单元可能只是碎片， 薄片染色的解释受多种伪影的影响。值得注意的是，IHC/IF可能无法证明 重要特征，如边缘挤压、淋巴细胞渗入或鳞状改变，为 诊断和治疗的不确定性。 组织光学透明和光学显微镜的最新进展使正常的3D荧光成像成为可能 不透明、厚的组织和器官。戏剧性的例子包括绘制整体神经元连接的进展图 啮齿动物的大脑处于亚细胞分辨率。我试图调整这些策略来推进炎症性疾病的分析 在整个小鼠肿瘤和人类肿瘤活检中的浸润物、药物输送和治疗反应。在……里面 为了推进3D肿瘤成像，我优化了光学透明、细胞染色、高分辨率成像和 计算重建。最终，我开发了一种3D肿瘤成像方法，透明肿瘤 体层摄影术(T3)，作为一种工具，以单细胞分辨率可视化和绘制肿瘤图。在这个项目中，我将应用T3 用于肿瘤免疫学和免疫治疗的空间分析。T3介导的3D肿瘤成像将提供 关于抗体药物分布和免疫环境的综合空间信息在整个肿瘤中 将作为一种新的检测工具用于癌症免疫治疗研究。

Project Summary This proposal aims to apply an imaging technique for three-dimensional (3D) visualization and mapping of the tumor and microenvironment including cancer cells, stromal cells, immune cells, and vascular cells. The capability to determine the location, density, and functional orientation of different cell populations throughout a tumor would be a powerful tool not only for cancer biology but also for improving diagnosis and advancing the development of effective therapies. Although immunohistochemistry (IHC)/immunofluorescence (IF) staining has long been used in cancer diagnostics, conventional IHC/IF methodology is based on localization of antigens in single thin sections. Considering that most cells in the field of view may be only fragments, interpretation of thin section staining is subject to multiple artifacts. Significantly, IHC/IF may fail to demonstrate important features such as pushing margin, lymphocytic infiltrate, or squamous change, leaving room for diagnostic and therapeutic uncertainties. Recent progress in tissue optical clearing and light microscopy enable 3D fluorescent imaging of normally opaque, thick tissues and organs. Dramatic examples include progress mapping neuronal connectivity in whole rodent brains at subcellular resolution. I sought to adapt these strategies to advance analysis of inflammatory infiltrates, drug delivery, and therapeutic responses in whole mouse tumors and human tumor biopsies. In order to advance 3D tumor imaging, I optimized optical clearing, cell staining, high resolution imaging, and computational reconstruction. Eventually, I developed a 3D tumor imaging method, Transparent Tumor Tomography (T3), as a tool to visualize and map tumors at single cell resolution. In this project, I will apply T3 for spatial analysis of tumor immunology and immunotherapy. T3-mediated 3D tumor imaging will provide integrated spatial information regarding antibody drug distribution and immune contexts in whole tumors which will be useful as a new assay tool for cancer immunotherapy research.