Imaging mechanisms of metastatic tumor formation in situ

原位转移性肿瘤形成的成像机制

• 批准号: 10374648
• 负责人: Gaudenz Danuser
• 金额: $ 168.85万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10374648
• 关键词: 3-Dimensional; Adaptor Signaling Protein; Address; Adopted; Affect; Algorithms; Architecture; Automobile Driving; Biological; Biological Assay; Biological Models; Blood Vessels; Cancer Biology; Cancer Etiology; Carcinoma in Situ; Cells; Cellular Morphology; Cellular biology; Cessation of life; Chemicals; Clinical Treatment; Communities; Computer Vision Systems; Core Facility; Data; Development; Disease; Distant; Educational workshop; Embryo; Environment; Environmental Risk Factor; Event; Ewings sarcoma; Extracellular Matrix; Goals; Heterogeneity; Human; Image; Imaging Device; Immunofluorescence Immunologic; In Situ; Individual; Informatics; Institution; Intelligence; Intrinsic factor; Invaded; Investigation; Label; Light; Malignant - descriptor; Malignant Bone Neoplasm; Malignant Neoplasms; Membrane; Membrane Proteins; Metabolic; Metabolism; Methods; Microscope; Microscopy; Molecular; Molecular Analysis; Molecular Probes; Morphology; Mus; NCI Center for Cancer Research; Neoplasm Metastasis; Oncogenic; Optics; Organ; Organism; Pattern; Periodicity; Physiological; Pilot Projects; Play; Primary Neoplasm; Process; Property; Research; Research Personnel; Resolution; Role; Sampling; Series; Side; Signal Transduction; Site; Solid Neoplasm; Specimen; Speed; Subgroup; System; Technology; Testing; Thick; Tissue imaging; Tissues; Training; Tropism; Variant; WNT Signaling Pathway; Work; Xenograft procedure; Zebrafish; cancer cell; caveolin 1; cell behavior; childhood sarcoma; cloud based; dimensional analysis; experimental study; fluorescence imaging; functional adaptation; high dimensionality; high resolution imaging; high throughput screening; imaging approach; imaging platform; imaging probe; imaging program; intravital imaging; lipid metabolism; live cell imaging; lymphatic vessel; melanoma; molecular imaging; multimodality; multiplexed imaging; novel; outreach; programs; quantitative imaging; response; technology development; tumor xenograft

Project Summary In response to the RFA for a Cellular Cancer Biology Imaging Program we propose a program focused on imaging and molecularly probing the cell biological events that drive the formation of new metastatic tumors. Specifically, we will address two questions: 1) How does the intersection of shifts in cell-intrinsic and cell- extrinsic signals associated with shifts in expression of the membrane adaptor protein Caveolin-1 affect the metastatic propensity of pediatric sarcoma (Research Testbed Unit 1)? 2) What are the effects of cell-intrinsic and cell-extrinsic variation in lipid metabolism on melanoma metastasis patterns (Research Testbed Unit 2)? Answers to both questions depend on technology to capture the molecular, metabolic, and morphological states of individual metastatic cells as they colonize the distant site: In the Technology Development Unit-1 we will develop a multi-modal, multi-scale live imaging platform to investigate the effects of intersecting microenvironmental variation across an organism and cell intrinsic heterogeneity on metastatic spreading. The platform will leverage the exquisite optical and physiological properties of the zebrafish embryos to ‘watch’ at once how cells form human tumor xenografts spread to multiple distant sites where they form metastatic tumors. The microscope will allow seamless switching between a high-throughput screening mode observing the metastatic patterns in tens to hundreds of embryos in one experiment and a high-resolution imaging mode with fully isotropic resolution of 300 nm in XYZ that allows detailed analysis of the molecular, metabolic, morphologic, and proliferation/survival states of individual cells within an emerging metastatic niche. In the Technology Development Unit-2 we will develop a multi-scale imaging platform to investigate by hyper-spectral analysis the molecular, metabolic, morphological, and functional states of metastatic cells across entire mouse organs. The platform will leverage advances in tissue clearing, fully automated high-speed and high-resolution light-sheet fluorescence imaging, and computer vision, to integrate a mesoscopic imaging mode for fast acquisition of volumes of up to 20 x 20 x 20 mm at a ~5-10 micron isotropic resolution with a nanoscopic imaging mode providing 300 nm XYZ-resolution throughout a 300 micron field of view anywhere in the organ. Biological features can thus be rapidly identified and immediately interrogated with high subcellular resolution. We will then develop physically and chemically accelerated 60-plex cyclic immunofluorescence assays to comprehensively characterize the molecular, metabolic and architectural states of colonizing cells and their surroundings in the metastatic niche in thick (~200 microns) tissue sections. To accurately describe metastatic heterogeneity, the entire system, including sample handling, labeling, and imaging, will be fully automated and operated in a high-throughput fashion. Our goal with this system is to enable comprehensive profiling of heterogeneous cell metastatic cell behavior in 100’s of intact tissue specimens. Together, these platforms will generate versatile imaging tools for a new era of in situ cancer cell biology.

项目摘要 为了响应用于细胞癌生物学成像计划的RFA，我们提出了一个针对的计划 成像和分子探测驱动新转移性肿瘤形成的细胞生物学事件。 具体而言，我们将解决两个问题：1）细胞中心和细胞转移的相交如何 与膜适配器蛋白可爱素1表达相关的外在信号会影响 小儿肉瘤的转移倾向（研究测试床单元1）？ 2）细胞中心的影响是什么 脂质代谢中的细胞 - 肢体变异对黑色素瘤转移模式（研究测试台2单元）？ 两个问题的答案都取决于捕获分子，代谢和形态学的技术 单个转移细胞在定居遥远位点时的状态：在技术开发中 将开发一个多模式，多尺度的实时成像平台，以研究相交的效果 在转移性扩散上的生物体和细胞固有异质性的微环境变异。 平台将利用斑马鱼胚胎的独家光学和物理特性来“手表” 一旦细胞形成人类肿瘤Xenographograptic如何扩展到多个远处形成转移性的位点 肿瘤。显微镜将允许在高通量筛选模式观察者之间进行无缝切换 在一个实验和高分辨率成像模式中，以数十万到数百个胚胎的转移模式的转移模式 在XYZ中的300 nm的完全各向同性分辨率可以详细分析分子，代谢， 新兴转移性生态位中各个细胞的形态和增殖/生存状态。在 技术开发单元2我们将开发一个多尺度成像平台，以通过超光谱调查 分析整个小鼠的转移细胞的分子，代谢，形态和功能状态 器官。该平台将利用组织清除，全自动高速和高分辨率的进步 荧光荧光成像和计算机视觉，以快速整合介观像模式 在〜5-10微米各向同性的分辨率上，以纳米镜分辨率获得高达20 x 20 x 20 mm的体积 成像模式在器官中任何地方的300微米视野中提供300 nm XYZ分辨率。 因此，可以快速识别生物学特征，并通过高细胞分辨率立即审问。 然后，我们将开发物理和化学加速的60粒环节免疫荧光测定到 全面地表征定植细胞及其的分子，代谢和建筑状态 厚（200微米）组织切片中转移性细分市场的周围环境。准确描述转移性 异质性，整个系统，包括样品处理，标签和成像，将完全自动化，并且 以高通量方式操作。我们使用该系统的目标是启动全面分析 在完整的组织标本中，异质细胞转移性细胞行为。这些平台将在一起 为原位癌细胞生物学的新时代生成多功能成像工具。