Hyperspectral and Structural Microscopy Platform for Therapy of Resistant Cancer

描述（由申请人提供）：尽管在检测和治疗癌症方面取得了很大进展，但播散性转移性疾病仍然是癌症患者死亡的主要原因。令人沮丧的是，由于这些致命病变的广泛性质，通常是亚临床大小，以及高度异质性的微环境，人们对这些致命病变如何在体内对治疗产生反应并最终抵抗治疗知之甚少。缺乏适当的成像工具来了解和克服转移性癌症在微观尺度上的治疗耐药性，这是癌症研究和治疗中一个主要的未满足的需求。为了解决这一关键挑战，本应用程序引入了一种创新的显微技术范式，用于可视化和理解弥漫性转移性癌症中发现的微尺度治疗反应和耐药因素。这项工作的目标是开发一个高通量、活细胞、分子和结构光学显微镜平台，能够在细胞和结节水平上观察和监测体外微转移病变的治疗反应。这样的成像平台将通过建立从单细胞到整个肿瘤的详细的癌症治疗时空图，为基础癌症和癌症治疗研究提供自下而上的方法。基于高光谱显微镜和光学相干断层扫描（OCT）的互补分子和结构成像技术，自动化多路治疗成像平台将实现长期的细胞和结节水平研究，在结构背景下绘制分子治疗因素的3D分布。新的实时，可调，近红外优化的高光谱显微镜系统将允许3D，多荧光团，肿瘤结节深部治疗抵抗因子的活细胞成像。为了对治疗引起的复杂结构变化进行纵向微观可视化，将与高光谱显微镜一起集成非摄动和无标记的延时OCT （TL-OCT）模式。适用于许多转移性癌症，这个新的成像平台将使用生理相关的、可重复的、多路复用的体外微转移性卵巢癌3D模型进行验证，用于基于成像的筛查。在这种方法的首次应用中，癌症生物学研究的多模式技术平台将专注于可视化和关联几个关键的治疗耐药因素，如药物摄取和扩散、缺氧和pH值与治疗反应。这种基于成像的方法系统和定量地表征癌症治疗反应和耐药性，与传统方法不同，传统方法错过了关键的分子和结构信息。这些技术可以很容易地转化为体外或原位显微内窥镜成像患者组织样本，用于图像引导的治疗计划。通过在微观尺度上建立我们对治疗反应和耐药性的知识，这个创新的成像平台形成了一个变革性的、由研究人员主导的新研究项目的基础，旨在改善当前的癌症治疗方法和击败治疗耐药的转移性疾病。

DESCRIPTION (provided by applicant): Although much progress has been made in detecting and treating cancer, disseminated metastatic disease remains the leading cause of death in cancer patients. Frustratingly little is known regarding how these deadly lesions respond to and eventually resist treatment in vivo due to their widespread nature, often sub-clinical sizes, and highly heterogeneous microenvironments. The lack of appropriate imaging tools to understand and overcome treatment resistance on the microscale in metastatic cancer represents a major unmet need in both cancer research and therapeutics. To address this critical challenge, this application introduces an innovative microscopic technological paradigm for visualizing and understanding the microscale treatment response and resistance factors found in disseminated metastatic cancer. The goal of this work is to develop a high- throughput, live-cell, molecular and structural optical microscopy platform capable of visualizing and monitoring therapeutic response in micrometastatic lesions in vitro at both the cellular and nodular levels. Such an imaging platform will enable a bottom-up approach for basic cancer and cancer therapeutics research by building a detailed spatiotemporal picture of cancer therapy from single cells all the way to the whole tumor. Based upon the complementary molecular and structural imaging technologies of hyperspectral microscopy and optical coherence tomography (OCT), the automated multiplexed therapy-imaging platform will enable long-term cellular and nodular-level studies that map the 3D distribution of molecular treatment factors within a structural context. The new real-time, tunable, near-IR optimized hyperspectral microscopy system will allow for 3D, multi-fluorophore, live-cell imaging of treatment resistance factors deep in tumor nodules. For longitudinal microscale visualization of the complex structural changes caused by treatment, the non- perturbative and label-free time-lapse OCT (TL-OCT) modality will be integrated along with the hyperspectral microscope. Applicable to many metastatic cancers, this new imaging platform will be validated using a physiologically relevant, repeatable, and multiplexed in vitro 3D model of micrometastatic ovarian cancer for imaging-based screens. In the first application of this approach, the multimodal technological platform for cancer biology research will focus on visualizing and correlating several crucial treatment resistance factors, such as drug uptake and diffusion, hypoxia, and pH, with treatment response. This imaging-based approach to systematically and quantitatively characterize cancer therapy response and resistance represents a departure from traditional methods that miss crucial molecular and structural information. These technologies can be readily translated for imaging patient tissue samples ex vivo, or in situ microendoscopically, for image-guided therapeutic planning. By building our knowledge of treatment response and resistance at the microscale, this innovative imaging platform forms the foundation of a transformative, new investigator-led research program aiming to improve current cancer therapeutics and defeat treatment-resistant metastatic disease. PUBLIC HEALTH RELEVANCE: Cancer that becomes metastatic is too often fatal due to the spread of numerous microscopic lesions that grow to be treatment resistant. The goal of this application is to address the root causes of treatment resistance in cancer through an innovative microscale imaging approach capable of visualizing the crucial molecular and structural factors involved in therapeutic response. By building an understanding of how deadly lesions escape therapy from the cellular level to the whole tumor, and using this knowledge to defeat treatment-resistant disease, the technology developed in this application aims to significantly improve the lives of patients suffering from advanced metastatic cancer.