Imaging mechanisms of metastatic tumor formation in situ

原位转移性肿瘤形成的成像机制

基本信息

批准号：
10491345
负责人：
Gaudenz Danuser
金额：
$ 160.93万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
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/10491345
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

项目摘要

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.

项目概要为了响应 RFA 的细胞癌症生物学成像计划，我们提出了一项重点关注的计划对驱动新转移性肿瘤形成的细胞生物学事件进行成像和分子探测。具体来说，我们将解决两个问题：1）细胞本征和细胞内转移的交集如何发生？与膜接头蛋白 Caveolin-1 表达变化相关的外在信号会影响儿童肉瘤的转移倾向（研究测试台单元 1） 2) 细胞内在的影响是什么？以及脂质代谢的细胞外在变化对黑色素瘤转移模式的影响（研究测试床单元 2）？这两个问题的答案取决于捕获分子、代谢和形态学特征的技术。单个转移细胞在远处定植时的状态：在技术开发单元 1 中，我们将开发一个多模态、多尺度的实时成像平台来研究相交的影响生物体的微环境变化和转移扩散的细胞内在异质性。平台将利用斑马鱼胚胎精致的光学和生理特性来“观看” 细胞如何形成人类肿瘤异种移植物扩散到多个遥远的部位，并在那里形成转移显微镜将允许在高通量筛选模式观察之间无缝切换。一次实验中数十至数百个胚胎的转移模式和高分辨率成像模式在 XYZ 中具有 300 nm 的完全各向同性分辨率，可以对分子、代谢、新兴转移生态位中单个细胞的形态和增殖/生存状态。技术开发单元2，我们将开发一个多尺度成像平台，通过高光谱进行研究分析整个小鼠转移细胞的分子、代谢、形态和功能状态该平台将利用组织透明化、全自动高速和高分辨率方面的进步。光片荧光成像和计算机视觉，集成介观成像模式以实现快速使用纳米显微镜以约 5-10 微米各向同性分辨率采集高达 20 x 20 x 20 毫米的体积成像模式可在器官内任意位置的 300 微米视野范围内提供 300 nm XYZ 分辨率。因此，可以快速识别生物特征并立即以高亚细胞分辨率进行询问。然后，我们将开发物理和化学加速的 60 重循环免疫荧光测定法，以全面表征定植细胞的分子、代谢和结构状态及其厚（约 200 微米）组织切片中转移生态位的周围环境，以准确描述转移。异质性，整个系统，包括样品处理、标记和成像，将完全自动化，我们使用该系统的目标是实现全面的分析。这些平台将共同研究数百个完整组织样本中的异质细胞转移细胞行为。为原位癌细胞生物学的新时代生成多功能成像工具。