New Ultrastructural 3D Optical Imaging of Tumor Endothelium for Cancer Nanomedicine Development

项目2：用于肿瘤纳米药物的新型肿瘤内皮超微结构三维光学成像 发展 摘要 该项目的长期目标是设计出更安全、更有效的新一代乳房 肿瘤纳米药物通过以下途径有效地克服血-肿瘤屏障而改善对肿瘤的药物输送 细胞穿透。作为第一步，我们提出了建立一种新的超结构3D超分辨率光学成像 以无标记方式跟踪和量化纳米颗粒在细胞内的运输和跨细胞作用的平台。 纳米粒转运是肿瘤纳米药物的一种新的传递途径，可通过两种主要途径发生 细胞内途径：(1)囊泡介导的细胞内转运；(2)纳米颗粒通过细胞内的穿梭 小管。然而，目前尚不清楚哪种途径会导致更有效的纳米颗粒转运，以及 到目前为止，这些路线倾向于特定的纳米颗粒尺寸。区分囊泡和小管介导的 纳米颗粒的传输途径，需要对整个内皮细胞进行超微结构的3D成像。我们的目标是 建立三维超分辨率光学显微镜作为了解细胞内的一种新的和独特的方法 纳米颗粒在细胞穿透过程中所采取的途径。使用我们独特的标签费3D超分辨率成像 方法，我们将检验纳米颗粒大小将决定纳米颗粒跨细胞途径的假设 和乳腺癌相关内皮细胞的跨细胞效率。为此，我们建议 遵循两个具体目标。目的1是建立乳腺癌的三维超分辨光学显微镜 相关内皮细胞与无标记纳米颗粒成像相结合。目标2是对纳米颗粒进行量化 人乳腺癌相关血管内皮细胞的转胞作用。为了以最佳方式进行建议的 研究任务和追求这个项目的成功，研究项目负责人(RPL)有两个经验 并组建了一支多学科研究团队，在 纳米医学、肿瘤血管生物学、超分辨率显微镜和乳腺肿瘤学。这个项目将 全面研究一种独特的成像方法，将其作为可应用于可视化的平台技术 并在超微结构分辨率下研究了纳米颗粒在不同组织和细胞中的传输 传统的光学显微镜。因此，使用这种新的成像方法可以帮助设计和实现 纳米粒子在细胞内传输和跨细胞传输的最佳性能显著 提高肿瘤组织给药在癌症治疗中的疗效。该项目的成功将为 支持RPL申请NIH R01项目的基本初步研究数据和科学证据(即， PAR-20-284-癌症纳米医学的创新研究)。

Project 2: New Ultrastructural 3D Optical Imaging of Tumor Endothelium for Cancer Nanomedicine Development ABSTRACT The long-term objective of this project is to engineer a new generation of safer and more effective breast cancer nanomedicines that improve drug delivery to tumors by efficiently overcoming the blood-tumor barrier via transcytosis. As a first step, we propose to establish a new ultrastructural 3D super-resolution optical imaging platform to track and quantify in a label-free manner the intracellular nanoparticle transport and transcytosis. Nanoparticle transcytosis is a novel delivery pathway in cancer nanomedicine and may occur through two major intracellular routes: (1) vesicle mediated intracellular transport, and (2) shuttling of nanoparticles via intracellular tubules. However, it is unknown which pathway results in more efficient nanoparticle transcytosis and whether these routes favor specific nanoparticle sizes to date. To differentiate between the vesicle and tubule-mediated nanoparticle transport routes, ultrastructural 3D imaging of whole endothelial cells is needed. Our objective is to establish a 3D super-resolution optical microscopy as a novel and unique method to understand the intracellular pathways that nanoparticles take during transcytosis. Using our unique label-fee 3D super-resolution imaging approach, we will test the hypothesis that nanoparticle size will determine the nanoparticle transcytosis pathway and transcytosis efficiency in breast-cancer associated endothelial cells. For this purpose, we propose the following two Specific Aims. Aim 1 is to establish 3D super-resolution optical microscopy of breast-cancer associated endothelial cells in combination with label-free nanoparticle imaging. Aim 2 is to quantify nanoparticle transcytosis in human breast cancer associated endothelial cells. In order to optimally conduct the proposed research tasks and pursue the success of this project, the research project leader (RPL) has two experienced mentors and also assembled a multidisciplinary research team with unique and synergistic expertise in the nanomedicine, tumor vascular biology, super-resolution microscopy, and breast oncology. This project will comprehensively investigate a unique imaging approach as a platform technology that can be applied to visualize and study the nanoparticle transport in different tissues and cells in 3D at ultrastructural resolution using conventional optical microscopes. As a result, using this novel imaging method can help design and implement the optimal nanoparticle properties for intracellular nanoparticle transport and transcytosis to significantly improve efficacy of the drug delivery to tumor tissues in cancer treatment. Success of this project will provide the essential preliminary study data and scientific evidence to support the RPL to apply for the NIH R01 project (i.e., PAR-20-284 – Innovative Research in Cancer Nanomedicine) in the future.