The rodent eye as a non-invasive window for understanding cancer nanotherapeutics

 描述（由申请人提供）：啮齿类动物的眼睛作为一个非侵入性的窗口，了解癌症nanotherapeutics摘要：我们建议使用小鼠眼睛作为一个非手术窗口，高效率，光学调查视网膜下异种移植模型和nanoderivery，使用国家的最先进的眼部成像仪，“EyePod”。EyePod采用单细胞分辨率活体共聚焦显微镜和光学相干断层扫描，通过眼睛的自然光学完全非侵入性地进行。该技术能够在数周甚至数月内进行可重复的体内成像，定量跟踪肿瘤发展和治疗诊断纳米颗粒的递送，以及测量肿瘤和组织反应。我们最近报道的新型胶束基纳米卟啉将用于该项目。这种令人兴奋的和高度通用的治疗诊断卟啉/胆酸为基础的 胶束纳米颗粒允许（i）有效包封疏水性化疗药物或荧光染料，（ii）通过卟啉的内源荧光进行肿瘤的近红外荧光（NIRF）检测，（iii）分别通过在肿瘤部位有效产生自由基和热进行光动力疗法（PDT）和光热疗法（PTT），（iv）用于MRI成像的Gd（III）负载，（v）用于PET成像的64 Cu负载，和（vi）将癌症靶向配体方便地连接到胶束表面用于癌症特异性靶向递送。深入了解这种纳米载体如何在肿瘤微环境中分布，以及它如何响应受控的光学刺激，将使我们能够最大限度地发挥其作为纳米治疗诊断剂的治疗潜力。我们最近还开发了一种短肽，它能被小鼠脑血管内皮细胞特异性吸收。具体目标是：1。在裸鼠眼中开发眼内胶质母细胞瘤和乳腺癌异种移植物模型，并使用非侵入性光学技术（EyePod）在细胞分辨率下纵向研究这些肿瘤模型在数天和数周内的发展，以及它们对纳米多柔比星治疗的反应。2.利用肿瘤模型和EyePod研究肿瘤靶向和非靶向纳米卟啉在肿瘤微环境中的体内生物分布和光响应，并利用冷冻电镜在超微结构水平上“解剖”纳米递送。3.优化脑血管内皮细胞穿透配体，并使用体内EyePod成像和冷冻电子显微镜研究其纳米递送到视网膜血管系统中，以及穿过与血脑屏障非常相似的血视网膜屏障。假设和影响：我们假设小鼠眼睛提供了一个独特而方便的窗口来研究：（i）肿瘤发展，（ii）肿瘤微环境内纳米颗粒药物或成像剂的生物分布，以及（iii）肿瘤对纳米治疗药物的反应的生物学。通过将先进的光学技术与最先进的无标记和荧光标记技术相结合，我们将能够在数天和数周的真实的时间内以微米分辨率检查纳米颗粒和封装药物的位置和命运。从这项研究中获得的信息不仅有助于理解纳米颗粒渗透到肿瘤血管系统中、在肿瘤基质中的分布以及摄取到肿瘤细胞中，而且还将有助于理解纳米颗粒在肿瘤血管系统中的分布。 使我们能够建立强大的协议，有效地研究肿瘤的发展，肿瘤微环境，体内肿瘤内的生物分布和命运的药物封装的纳米载体，和纳米输送通过BRB。

 DESCRIPTION (provided by applicant): The rodent eye as a non-invasive window for understanding cancer nanotherapeutics Abstract: We propose to use the mouse eye as a non-surgical window for highly efficient, optical investigation of subretinal xenograft models and nanodelivery, using a state-of-the-art ocular imager, "EyePod". The EyePod employs single-cell resolution intravital confocal microscopy and optical coherence tomography, performed completely non-invasively through the natural optics of the eye. This technology enables repeatable in vivo imaging over weeks and even months, quantitative tracking of tumor development and delivery of theranostic nanoparticles, and the measurement of tumor and tissue responses. The novel micellar-based nanoporphyrin that we have recently reported will be used in this project. This exciting and highly versatile theranostic porphyrin/cholic acid-based micellar nanoparticle allows (i) efficient encapsulation of hydrophobic chemotherapeutic drugs or fluorescent dyes, (ii) near- infra red fluorescent (NIRF) detection of the tumor via the intrinsc fluorescence of porphyrins, (iii) photodynamic therapy (PDT) and photothermal therapy (PTT) via efficient free radical and heat generation at the tumor site, respectively, (iv) Gd(III) loadin for MRI imaging, (v) 64Cu loading for PET imaging, and (vi) convenient ligation of cancer-targeting ligands to the surface of the micelle for cancer- specific targeted delivery. Thorough understanding of how this nanocarrier distributes within the tumor microenvironment, and how it responds to controlled optical stimulation will enable us to maximize its therapeutic potential as a nano-theranostic agent. We have also recently developed a short peptide that is specifically taken up by mouse brain vascular endothelial cells. Specific Aims are: 1. To develop intraocular glioblastoma and breast cancer xenograft models in eyes of nude mice, and to use non-invasive optical techniques (the EyePod) to study the development of these tumor models at cellular resolution longitudinally over days and weeks, and their response to treatment with nanodoxorubicin. 2. To use the tumor models and EyePod to study the biodistribution and photo-response of tumor targeting and non-targeting nanoporphyrins within the tumor micro-environment in vivo, and to use cryo-electron microscopy to "dissect" the nanodelivery at the ultrastructural level. 3. To optimize brain vascular endothelial cell-penetrating ligands, and to use in vivo EyePod imaging and cryo-electron microscopy to study their nanodelivery into retinal vasculature, and across the blood retinal barrier, which is very similar to blood brain barrier. Hypothesis & Impact: We hypothesize that the mouse eye provides a unique and convenient window to study: (i) tumor development, (ii) biodistribution of nanoparticle drugs or imaging agents inside the tumor microenvironment, and (iii) biology of tumor responses to the nanotherapeutics. By combining advanced optics with state-of-the art label-free and fluorescence labeling, we will be able to examine at micrometer resolution the location and fate of the nanoparticles and the encapsulated drug in real time over days and weeks. Information gained from this study will not only facilitate understanding of nanoparticle penetration into the tumor vasculature, distribution inside the tumor stroma, and uptake into tumor cells, but will also allow us to establish robust protocols for efficient study of tumor development, tumor microenvironment, in vivo intra-tumoral biodistribution and fate of drugs encapsulated by nanocarrier, and nanodelivery across the BRB.