In Vivo Molecular Imaging of the Retina

描述（由申请人提供）：通过仪器的进步，动物模型和患者视网膜成像技术得到了显著改善，实现了组织结构的高分辨率成像。此外，数十年的基础和临床研究已经确定了许多分子生物标志物，这些标志物可能对评估视网膜疾病的易感性、早期疾病和疾病进展以及临床前研究中的分子机制有价值。本提案的目标是建立在成像仪器和生物标志物研究的进步基础上，以开发视网膜体内分子成像技术。该策略基于发夹功能化金纳米颗粒（hAuNP），这是一种生物相容性金胶体，经过工程设计，可以进入活组织，并在与目标信使RNA （mRNA）或microRNA序列杂交后发出荧光。最近发表的研究和初步研究表明，hAuNP能够特异性靶向哺乳动物细胞和视网膜脉管系统中的多种不同RNA序列，而不会对细胞功能产生不良影响。在这项提议中，hAuNP将利用激光诱导脉络膜新生血管（LCNV）的小鼠模型来验证mrna和microrna作为脉络膜新生血管（CNV）的生物标志物。在Aim 1中，hAuNP将用于对原代脉络膜内皮细胞和视网膜色素上皮细胞中cnv相关生物标志物进行成像，hAuNP的生物分布和安全性将在小鼠模型中进一步测试。在该提案的目标2和3中，hAuNP将在LCNV小鼠模型中进行评估，以建立纵向，多路RNA成像的效用。这些研究将为RNA和其他分子生物标志物在动物模型中的分子成像建立框架，并将促进这些技术在患者疾病早期检测和分期方面的临床转化。

DESCRIPTION (provided by applicant): Technologies for imaging the retina in animal models and patients have been significantly improved through advances in instrumentation, enabling high-resolution imaging of tissue structure. Furthermore, decades of basic and clinical research have identified a number of molecular biomarkers which may be valuable for assessing susceptibility to retinal disease, early disease, and disease progression, as well as dissecting molecular mechanisms in preclinical studies. The goal of this proposal is to build upon advances in imaging instrumentation and biomarker research in order to develop technologies for in vivo molecular imaging of the retina. The strategy is based on hairpin functionalized gold nanoparticles (hAuNP), biocompatible gold colloids engineered to enter living tissues and fluoresce upon hybridization with targeted messenger RNA (mRNA) or microRNA sequences. Recently published and preliminary studies demonstrate that hAuNP are capable of specifically targeting multiple distinct RNA sequences in mammalian cells and the retinal vasculature, without adverse effects on cell function. In this proposal, hAuNP will be utilized to validate mRNAs and microRNAs as biomarkers of choroidal neovascularization (CNV), using a mouse model of laser- induced choroidal neovascularization (LCNV). In Aim 1, hAuNP will be used to image CNV-relevant biomarkers in primary choroidal endothelial cells and retinal pigment epithelial cells, and the biodistribution and safety profiles of hAuNP will be further tested in mouse models. In Aims 2 and 3 of the proposal, hAuNP will be evaluated in a mouse model of LCNV in order to establish the utility of longitudinal, multiplexed RNA imaging. These studies will set the framework for molecular imaging of RNA and other molecular biomarkers in animal models, and will facilitate clinical translation of these technologies for early detection and staging of disease in patients.