A Portable low-cost, Point of Investigation CapCell Scope to Image and Quantify the Major Axes of Metabolism and the Associated Vasculature in In vitro and In vivo Biological Models

摘要 对新陈代谢和血管微环境的兴趣在广泛的学科中继续增长 包括神经科学、心血管生物学和癌症研究领域。鉴于它们在临床上的重要性， 令人惊讶的是，几乎没有可用的技术可以在系统级别上一起提供这些特征的视图 活着。尽管有许多台式显微镜和代谢工具可以提供精致的分辨率 而对于新陈代谢或血管成像，它们通常需要研究人员将动物运送到专门的 设施，这限制了对高频成像的访问，这些系统需要广泛的培训，并且经常 视场(FOV)、分辨率和波长，仅适用于最常见的使用情形(在光学情况下 显微镜系统)。我们的目标是开发和验证研究点代谢和血管成像 系统和相应算法将使个人对癌症研究的成像访问大众化 癌症药理实验室。我们系统的能力将通过新陈代谢成像来展示， 血管功能和建筑的空间分辨率，可以可视化的增殖，退化，残留和 乳腺癌模型中的复发性疾病。我们的技术可以提供一个全面的理解 肿瘤的代谢特征影响治疗结果，可用于多种癌症 应用药理学来提高癌症治疗的有效性。这项提议的具体目标是 开发低成本、便携、用户友好的CapCell显微镜，以成像关键的代谢和血管终点 同时具有广泛的视场和分辨率(目标1)，证明该技术提供 在已建立的小鼠肿瘤模型中提供可比较的信息并可以补充已建立的方法 乳腺癌(Aim 2)，并在GEM模型来源的乳腺和活体乳腺上验证技术 GEM模型衍生肿瘤的窗口模型(目标3)。这项提议为将我们的 患者来源的异种移植(PDX)模型和患者来源的有机物的技术，已被证明是 能够真实地概括患者肿瘤的许多微环境特征，使我们能够移动我们的 技术向转化型药学研究迈进。

Summary Interest in metabolism and the vascular microenvironment continues to grow in a broad range of disciplines including neuroscience, cardiovascular biology, and the field of cancer research. Given their clinical importance, there are surprisingly few techniques available that can provide a systems-level view of these hallmarks together in vivo. Though there are many bench-top microscopes and metabolic tools available to provide exquisite resolution and contrast for metabolic or vascular imaging, they often require researchers to transport animals to specialized facilities and this limits access for high frequency imaging, these systems require extensive training and often have fields of view (FOV), resolution and wavelengths that fits only the most common use cases (in the case of optical microscopy systems). Our goals are to develop and validate a point-of-investigation metabolic and vascular imaging system and corresponding algorithms that will democratize access to imaging for cancer research in individual cancer pharmacology labs. The capabilities of our system will be demonstrated through imaging of metabolism, vascular function, and architecture at a spatial resolution that can visualize proliferative, regressive, residual, and recurrent disease in a breast cancer model. Our technique could provide a comprehensive understanding how the metabolic characteristics of tumors impact therapeutic outcome and can be used in a wide range of cancer pharmacology applications to improve the effectiveness of cancer therapy. The specific goals of this proposal are to develop an low-cost, portable, user friendly, CapCell scope to image key metabolic and vascular endpoints simultaneously with a wide range of FOVs and resolutions (Aim 1), demonstrate that this technology provides comparable information and can complement established methods in well-established murine tumor models of breast cancer (Aim 2), and validate the technology on GEM model derived mammospheres and in vivo mammary window models of GEM model derived tumors (Aim 3). This proposal sets the foundation for translating our technology to patient-derived xenograft (PDX) models and patient-derived organoids, which have been shown to be able to faithfully recapitulate many of the micro-environmental features of patient tumors, allowing us move our technique forward towards translational pharmaceutical research.