MAGNETIC NANOSENSORS FOR BIOMEDICAL ANALYSES OF MICROVESICLES

描述（由申请人提供）：微泡（MV）是由细胞释放到循环中的磷脂囊泡，最近已成为新的诊断生物标志物。在各种恶性肿瘤中，包括心血管疾病、糖尿病和炎症，已经报道了MV水平升高; MV似乎也在红细胞老化过程中发挥不可或缺的作用。然而，一个主要的障碍，以提高我们的知识MV生物学，从而充分利用其临床潜力，一直缺乏准确和标准化的方法MV分析。我们最近开发了一种新的基于纳米技术的诊断平台，称为“DMR”（诊断磁共振）。通过采用核磁共振（NMR）的原理，DMR设备测量样品的横向弛豫，从而用分子特异性磁性纳米颗粒（MNP）标记生物靶标。通过系统地开发优化的MNP和基于芯片的微型NMR系统，DMR技术现在已经取得了显着进步，可以提供灵敏的细胞即时分子分析。在这些成就的基础上，本提案的总体目标是调整并进一步推进DMR平台，以直接从全血中快速检测和多重分析MV。我们将特别关注以下方面 目标。在目标1中，我们将合成新的磁性纳米剂和测定方法，这将允许MV靶点的高效和选择性MNP标记。在目标2中，我们将实现一个小型化的NMR系统与复杂的微流体集成。该系统将被设计为使MV分析能够完全在单个芯片上进行;它将直接从全血中分离MV，用MNP标记MV，并对靶向MV进行NMR测量。在目标3中，优化的纳米试剂和装置都将应用于血液制品中红细胞衍生MV的检测和综合分析。这项研究将促进我们对血液老化生物学的理解，从而提高血液制品质量和输血安全性。我们设想拟议的DMR-MV技术在生命科学和临床实践中具有广泛的诊断潜力。通过促进来自不同细胞来源的MV的快速和定量分子分析，该技术可以实现早期疾病检测和治疗监测。反过来，这可以通过提供关于个体患者的细胞/分子特征的有价值的信息来加快创建个性化治疗的进展。 公共卫生相关性：我们建议开发一种新的基于纳米技术的平台，用于高度敏感的医疗诊断。该研究是高度协同的，整合了磁性纳米材料，新型生物共轭化学技术和微电子学的优势。开发的平台将用于检测和筛选血液中的微泡，这是血管疾病和输血医学中的紧急诊断生物标志物。

DESCRIPTION (provided by applicant): Microvesicles (MVs) are phospholipid vesicles released into the circulation by cells, which have recently emerged as a new diagnostic biomarker. Elevated level of MVs has been reported in various malignancies, including cardiovascular diseases, diabetes, and inflammation; MVs also appear to play an integral role in the erythrocyte aging process. However, a major barrier to advancing our knowledge of MV biology, and thus fully harnessing their clinical potential, has been the lack of accurate and standardized methods for MV analysis. We have recently developed a new, nanotechnology-based diagnostic platform termed "DMR" (diagnostic magnetic resonance). By employing principles of nuclear magnetic resonance (NMR), the DMR device measures the transverse relaxation of samples, whereby biological targets are labeled with molecular-specific magnetic nanoparticles (MNPs). By systematically developing optimized MNPs and chip-based miniature NMR systems, the DMR technology has now significantly advanced so as to provide sensitive, point-of-care molecular analyses of cells. Building upon these achievements, the overall goal of this proposal is to adapt and further advance the DMR platform for rapid detection and multiplexed profiling of MVs directly from whole blood. We will specifically focus on the following aims. In Aim 1, we will synthesize new magnetic nanoagents and assay methods that will allow highly efficient and selective MNP-labeling of MV targets. In Aim 2, we will implement a miniaturized NMR system integrated with sophisticated microfluidics. This system will be designed to enable MV analysis to be performed entirely on a single chip; it will isolate MVs directly from whole blood, label MVs with MNPs, and perform NMR measurements on the targeted MVs. In Aim 3, both the optimized nanoagents and device will be applied to the detection and comprehensive profiling of erythrocyte-derived MVs in blood products. This study will advance our understanding of the biology of blood aging, which could lead to improved blood product quality and transfusion safety. We envision a broad diagnostic potential for the proposed DMR-MV technology in both the life sciences and in clinical practice. By facilitating the rapid and quantitative molecular analysis of MVs from different cellular origins, this technology could enable early disease detection and treatment monitoring. In turn, this could expedite advances in creating personalized treatment by providing valuable information on the cellular/molecular signatures of individual patients. PUBLIC HEALTH RELEVANCE: We propose to develop a new, nanotechnology-based platform for highly sensitive medical diagnosis. The research is highly synergistic, integrating advantages of magnetic nanomaterials, novel bioconjugation chemistry technology, and microelectronics. The developed platform will be applied to detect and screen microvesicles in blood, an emergent diagnostic biomarker in vascular diseases and transfusion medicine.