In Vivo Molecular Imaging of the Retina

描述(由申请人提供)：通过仪器的进步，在动物模型和患者中对视网膜进行成像的技术已经得到显著改进，使得能够对组织结构进行高分辨率成像。此外，几十年的基础和临床研究已经确定了一些分子生物标志物，它们可能对评估视网膜疾病、早期疾病和疾病进展的易感性以及在临床前研究中剖析分子机制有价值。这项提议的目标是建立在成像仪器和生物标记物研究的进展基础上，以开发视网膜的活体分子成像技术。该策略基于发夹功能化的金纳米颗粒(HAuNP)，这是一种生物相容的金胶体，经过设计可以进入活组织，并在与靶向信使RNA(MRNA)或microRNA序列杂交时发出荧光。最近发表的初步研究表明，hAuNP能够特异性地靶向哺乳动物细胞和视网膜血管系统中的多种不同的RNA序列，而不会对细胞功能产生不利影响。在这项建议中，hAuNP将用于验证mRNAs和microRNAs作为脉络膜新生血管(CNV)的生物标志物，使用激光诱导的脉络膜新生血管(LCNV)的小鼠模型。在目标1中，hAuNP将用于原代脉络膜内皮细胞和视网膜色素上皮细胞中CNV相关生物标志物的成像，并将在小鼠模型中进一步测试hAuNP的生物分布和安全性。在该提案的目标2和3中，将在LCNV的小鼠模型中评估hAuNP，以建立纵向、多路复用的RNA成像的实用性。这些研究将为在动物模型中进行RNA和其他分子生物标记物的分子成像奠定基础，并将促进这些技术的临床转化，以便对患者的疾病进行早期检测和分期。

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.