Nanomedicine Lab, Faculty of Biology, Medicine & Health, AV Hill Building, The University of Manchester, Manchester, M13 9PT, United Kingdom.
Division of Cell Matrix Biology & Regenerative Medicine, Faculty of Biology, Medicine & Health, Biological, The University of Manchester, Manchester, M13 9PT, United Kingdom.
Biomaterials. 2019 Jan;188:118-129. doi: 10.1016/j.biomaterials.2018.10.011. Epub 2018 Oct 13.
The prominent discrepancy between the significant investment towards plasma biomarker discovery and the very low number of biomarkers currently in clinical use stresses the need for discovery technologies. The discovery of protein biomarkers present in human blood by proteomics is tremendously challenging, owing to the large dynamic concentration range of blood proteins. Here, we describe the use of blood-circulating lipid-based nanoparticles (NPs) as a scavenging tool to comprehensively analyse the blood proteome. We aimed to exploit the spontaneous interaction of NPs with plasma proteins once injected in the bloodstream, known as 'protein corona', in order to facilitate the capture of tumor-specific molecules. We employed two different tumor models, a subcutaneous melanoma model (B16-F10) and human lung carcinoma xenograft model (A549) and comprehensively compared by mass spectrometry the in vivo protein coronas formed onto clinically used liposomes, intravenously administered in healthy and tumor-bearing mice. The results obtained demonstrated that blood-circulating liposomes surface-capture and amplify a wide range of different proteins including low molecular weight (MW) and low abundant tumor specific proteins (intracellular products of tissue leakage) that could not be detected by plasma analysis, performed in comparison. Most strikingly, the NP (liposomal) corona formed in the xenograft model was found to consist of murine host response proteins, as well as human proteins released from the inoculated and growing human cancer cells. This study offers direct evidence that the in vivo NP protein corona could be deemed as a valuable tool to enrich the blood proteomic analysis and to allow the discovery of potential biomarkers in experimental disease models.
在投入大量资金用于发现血浆生物标志物的情况下，目前仅有极少数生物标志物可用于临床，这两者之间存在显著差异，凸显出对发现技术的需求。由于血液蛋白的动态浓度范围较大，通过蛋白质组学发现人类血液中存在的蛋白质生物标志物极具挑战性。在这里，我们描述了使用基于血液中环脂质的纳米颗粒 (NPs) 作为一种清除工具，全面分析血液蛋白质组。我们旨在利用 NPs 与注入血流后与血浆蛋白自发相互作用（称为“蛋白质冠”），以便于捕获肿瘤特异性分子。我们使用了两种不同的肿瘤模型，皮下黑色素瘤模型（B16-F10）和人肺癌异种移植模型（A549），并通过质谱法全面比较了临床应用的脂质体在健康和荷瘤小鼠体内形成的体内蛋白质冠。结果表明，血液循环中的脂质体表面捕获并放大了广泛的不同蛋白质，包括低分子量（MW）和低丰度的肿瘤特异性蛋白质（组织渗漏的细胞内产物），这些蛋白质无法通过比较进行的血浆分析检测到。最引人注目的是，在异种移植模型中形成的 NP（脂质体）冠包含了鼠宿主反应蛋白以及从接种和生长的人癌细胞中释放的人蛋白。这项研究提供了直接证据，表明体内 NP 蛋白质冠可以被视为一种有价值的工具，用于丰富血液蛋白质组学分析并发现实验疾病模型中的潜在生物标志物。
