Hyperspectral and Structural Microscopy Platform for Therapy of Resistant Cancer

描述(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，往往是亚临床大小，以及高度异质性的微环境，人们对这些致命病变在体内如何反应并最终抵抗治疗知之甚少。在转移性癌症中，缺乏适当的成像工具来了解和克服微观层面的治疗阻力，是癌症研究和治疗中尚未得到满足的主要需求。为了解决这一关键挑战，该应用程序引入了一种创新的微观技术范例，用于可视化和了解在播散性转移癌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、分子和结构的光学显微镜平台，能够在细胞和结节水平上可视化和监测体外微转移灶的治疗反应。这样的成像平台将通过构建从单个细胞到整个肿瘤的癌症治疗的详细时空图像，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成像(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能够进行长期的细胞和结节水平的研究，绘制出分子治疗因素在结构背景下的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对肿瘤结节深处的治疗耐药因素进行3D、多荧光团、活细胞成像。对于治疗引起的复杂结构变化的纵向微观可视化，非微扰和无标记的时延OCT(TL-OCT)模式将与高光谱显微镜集成在一起。这一新的成像平台适用于许多转移性癌症，将使用生理上相关的、可重复的和多路传输的微转移卵巢癌体外3D模型进行验证，以进行基于成像的筛查。在这一方法的首次应用中，癌症生物学研究的多模式技术平台将专注于可视化并将药物摄取和扩散、低氧和pH等几个关键的治疗耐药因素与治疗反应相关联。这种基于成像的方法系统地和定量地描述癌症治疗的反应和耐药性，与传统方法背道而驰，因为传统方法缺少关键的分子和结构信息。这些技术可以很容易地转化为患者组织样本的体外成像，或在原位显微内窥镜下，用于图像引导的治疗计划。通过在微观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、由研究人员领导的新研究计划的基础，该计划旨在改进当前的癌症治疗方法，并击败耐药转移疾病。

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.