State-of-the-art biological nanoparticle tracking instrument for interdisciplinary use

最先进的跨学科生物纳米粒子追踪仪器

• 批准号: RTI-2020-00372
• 负责人: Vijayan, Mathilakath
• 金额: $ 10.91万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693677
• 关键词: State; art; biological; nanoparticle; tracking

Most cells release small membrane-bound vesicles (30-100 nm) termed exosomes, a subpopulation of extracellular vesicles (EVs). Exosomes contain nucleic acids and proteins from the intercellular milieu and will interact with target cells to release a payload, which has the potential to stimulate secondary signalling cascades, including gene transcription. Consequently, exosomes have the potential to enact profound biological effects on the target cells, but the impact of this inter-tissue communication on the whole organism is unclear. From a therapeutics perspective, EVs have become increasingly important as biomarkers of diseases, such as cancers. EVs have also garnered interest as a drug delivery tool, as these vesicles are known to specifically target certain tissues. The extraordinary importance of these biologically active vesicles cannot be overstated; however, due to their small size, identifying and quantifying EVs in vivo has proved challenging. The University of Calgary has a thriving exosome research contingent, spanning three departments (Biological Sciences, Cumming School of Medicine, and Veterinary Medicine), and has resulted in many publications. A noted limitation in all these studies was the inability to quantify EV/exosome concentration in biological fluids. This not only curtails the current research capabilities, but also prevents the research from being competitive on the international stage. Nanoparticle tracking analysis (NTA) is a widely used method of quantifying both size and concentration of EVs in a dilute solution, and this technology is the gold standard for exosome research. Additionally, the NTA method can integrate fluorescent detection, allowing the ability to identify subpopulations of exosomes in a single sample. Particle Metrix's Zetaview TWIN uses NTA technology to detect vesicle size and concentration but additionally will detect two different fluorophores. This is a state-of-the-art instrument capable of measuring vesicles from the nanometer to micrometer size, and would be instrumental in producing high-quality, high-throughput data. Overall, this instrument is a necessary addition to support existing infrastructure in place at UC to study exosomal transport. In particular, ZetaView TWIN is necessary to quantify EVs/exosomes from in vivo studies. For example, does a drug treatment change the size, concentration or particular subpopulation of fluorescently tagged exosomes? Will stress or toxicant exposure in wild and domesticated fishes result in changes to the plasma exosome profile? Will the exosome released by microbes influence the gut epithelial cells? These are just a few of the questions that the primary applicant and co-applicants are hoping to answer using the Zetaview TWIN. The acquisition of the equipment will be critical for the next phase of EV research at the University of Calgary, and will support high-quality HQP recruitment and training, as well as facitlitating breakthrough research. **

大多数细胞释放称为外泌体的小膜结合囊泡（30-100 nm），它是细胞外囊泡（EV）的一个亚群。外泌体含有来自细胞间环境的核酸和蛋白质，并将与靶细胞相互作用以释放有效负载，该有效负载有可能刺激次级信号级联，包括基因转录。因此，外泌体有可能对靶细胞产生深远的生物学效应，但这种组织间通讯对整个生物体的影响尚不清楚。从治疗的角度来看，EV 作为癌症等疾病的生物标志物变得越来越重要。 EV 作为药物输送工具也引起了人们的兴趣，因为已知这些囊泡专门针对某些组织。这些具有生物活性的囊泡的非凡重要性怎么强调也不为过。然而，由于电动汽车尺寸较小，在体内识别和量化电动汽车已被证明具有挑战性。卡尔加里大学拥有一支蓬勃发展的外泌体研究队伍，横跨三个系（生物科学、卡明医学院和兽医），并发表了许多出版物。所有这些研究的一个值得注意的局限性是无法量化生物体液中的 EV/外泌体浓度。这不仅削弱了现有的研究能力，也阻碍了研究在国际舞台上的竞争力。纳米颗粒跟踪分析 (NTA) 是一种广泛使用的量化稀溶液中 EV 大小和浓度的方法，该技术是外泌体研究的黄金标准。此外，NTA 方法可以集成荧光检测，从而能够识别单个样品中的外泌体亚群。 Particle Metrix 的 Zetaview TWIN 使用 NTA 技术来检测囊泡大小和浓度，此外还可以检测两种不同的荧光团。这是一种最先进的仪器，能够测量从纳米到微米尺寸的囊泡，并将有助于产生高质量、高通量的数据。总的来说，该仪器是支持加州大学现有基础设施研究外泌体运输的必要补充。特别是，ZetaView TWIN 对于量化体内研究中的 EV/外泌体是必要的。例如，药物治疗是否会改变荧光标记外泌体的大小、浓度或特定亚群？野生和驯养鱼类的压力或有毒物质暴露是否会导致血浆外泌体谱发生变化？微生物释放的外泌体会影响肠道上皮细胞吗？这些只是主申请人和共同申请人希望使用 Zetaview TWIN 回答的一些问题。该设备的采购对于卡尔加里大学下一阶段的电动汽车研究至关重要，并将支持高质量的总部人员招募和培训，并促进突破性研究。 **