An intra-vital metabolic microscope to reveal the mechanisms of radiation resistance in head and neck carcinomas

项目摘要 鉴于放射抗性在头颈部鳞状细胞癌（HNSCC）中的临床重要性， 了解抗辐射肿瘤如何重新连接其代谢途径和血管网络， 放射治疗（RT）对于开发消除残留肿瘤细胞和/或预防肿瘤复发的策略至关重要。 随后复发。目前，没有技术可用于提供系统级方法来对图像进行成像。 代谢的主轴和相关的脉管系统在空间分辨率，可以阐明调制 癌细胞代谢或体内血管重编程。我们的技术目标是创造创新 显微镜解决方案，自动算法和实验策略，以成像肿瘤代谢，血管 在空间分辨率下的功能和结构，允许原发性肿瘤、残留疾病 和RT后复发，以促进对肿瘤生物学和功能的理解， 复发风险和治疗设计，以减轻残留疾病和/或在临床前完全复发 模型我们的技术方法填补了体外细胞研究和整体细胞研究之间存在的一个重要空白。 身体成像，并补充代谢组学和免疫组织化学。具体目标是 建议开发一种结合结构照明显微镜的便携式多参数显微镜 和暗场显微镜在再发射几何成像的关键代谢和血管终点 同时（具体目标1）;并使用体内HNSCC正交各向异性模型的技术来测试 一种新的假说，即RT诱导的缺氧诱导因子（HIF-1α和HIF-2α）表达和随后的缺氧诱导因子（HIF-1α和HIF-2α）表达可能是一种新的机制。 代谢/脉管系统的变化是HNSCC放射抗性的基础（具体目标2）。这一提议将使 将我们的技术转化为患者来源的异种移植模型的基础， 忠实地概括了患者肿瘤的许多微环境特征，使我们能够移动我们的 技术向转化药物研究迈进。

PROJECT SUMMARY Given the clinical importance of radio-resistance in head and neck squamous cell cancer (HNSCC), understanding how radio-resistant tumors rewire their metabolic pathways and vascular network to escape radiotherapy (RT) is critical towards developing strategies to eliminate residual tumor cells and/or prevent subsequent recurrence. Currently, no techniques are available to provide a systems level approach to image the major axes of metabolism and the associated vasculature at a spatial resolution that can elucidate the modulation of cancer cell metabolism or vascular reprogramming in vivo. Our technological goal is to create innovative solutions in microscopy, automated algorithms and experimental strategies to image tumor metabolism, vascular function and architecture at a spatial resolution that allows for visualization of primary tumors, residual disease and recurrence following RT to facilitate the understanding of tumor biology and function, assessment of recurrence risk and design of therapies to mitigate residual disease and/or recurrence altogether in pre-clinical models. Our technological approach fills an important gap that exists between in vitro cell studies and whole- body imaging, and is complementary to metabolomics and immunohistochemistry. The Specific Aims of this proposal are to develop a portable multi-parametric microscope that combines structured illumination microscopy and dark field microscopy in a re-emission geometry to image key metabolic and vascular endpoints simultaneously (Specific Aim 1); and use the technology with in vivo HNSCC orthotropic models to test the novel hypothesis that RT-induced hypoxia-inducible factors (HIF-1α and HIF-2α) expression and subsequent changes in metabolism/vasculature underlie HNSCC radio-resistance (Specific Aim 2). This proposal will set the foundation for translating our technology to patient-derived xenograft models that have been shown to faithfully recapitulate many of the micro-environmental features of patient tumors, allowing us to move our technique forward towards translational pharmaceutical research.