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

项目2：用于癌症纳米医学的肿瘤内皮细胞的新型超微结构3D光学成像 发展 摘要 该项目的长期目标是设计新一代更安全，更有效的乳房 癌症纳米药物，通过有效地克服血液-肿瘤屏障， 转胞吞作用作为第一步，我们建议建立一种新的超微结构三维超分辨率光学成像 该平台以无标记的方式跟踪和定量细胞内纳米颗粒转运和转胞吞作用。 纳米颗粒转胞吞作用是癌症纳米医学中的一种新的递送途径， 细胞内途径：（1）囊泡介导的细胞内转运，和（2）纳米颗粒经由细胞内途径的穿梭 小管然而，尚不清楚哪种途径导致更有效的纳米颗粒转胞吞作用，以及是否 迄今为止，这些途径有利于特定的纳米颗粒尺寸。为了区分囊泡介导的和小管介导的 为了确定纳米颗粒的运输途径，需要对整个内皮细胞进行超微结构3D成像。我们的目标是 建立一个三维超分辨率光学显微镜作为一种新颖的和独特的方法来了解细胞内 纳米颗粒在转胞吞作用中所采取的途径。使用我们独特的标签费3D超分辨率成像 方法，我们将测试假设，纳米颗粒的大小将决定纳米颗粒转胞吞途径 和乳腺癌相关内皮细胞中的转胞吞作用效率。为此，我们建议 有两个具体目标。目的1建立乳腺癌三维超分辨光学显微镜 相关的内皮细胞与无标记纳米颗粒成像的组合。目的2是量化纳米颗粒 在人乳腺癌相关内皮细胞中的转胞吞作用。为了最佳地进行拟议的 研究任务和追求这个项目的成功，研究项目负责人（RPL）有两个经验丰富的 导师，还组建了一个多学科的研究团队，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。该项目将 全面研究一种独特的成像方法，作为一种平台技术，可以应用于可视化 并研究纳米颗粒在不同组织和细胞中的三维超微结构分辨率， 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 用于细胞内纳米颗粒转运和转胞吞作用的最佳纳米颗粒性质显著 提高癌症治疗中药物递送至肿瘤组织的功效。该项目的成功将提供 支持RPL申请NIH R 01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.