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. **

大多数细胞释放出小膜结合的囊泡（30-100 nm），称为外泌体，细胞外囊泡的亚群（EV）。外泌体包含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载荷，有效载荷有可能刺激二级信号级联反应，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作用，但是这种组织间交流对整个生物体的影响尚不清楚。从治疗学的角度来看，随着癌症等疾病的生物标志物，电动汽车变得越来越重要。电动汽车还引起了作为药物输送工具的兴趣，因为已知这些囊泡专门针对某些组织。这些生物活性囊泡的非凡重要性不能被夸大。但是，由于它们的体积较小，因此在体内识别和量化电动汽车已被证明是具有挑战性的。卡尔加里大学有一个繁荣的外部研究特遣队，跨越了三个系（生物科学，卡明医学院和兽医医学），并导致了许​​多出版物。在所有这些研究中，一个著名的局限性是无法量化生物流体中的EV/外泌体浓度。这不仅削弱了当前的研究能力，而且还阻止了研究在国际舞台上具有竞争力。纳米颗粒跟踪分析（NTA）是一种量化稀释溶液中的大小和浓度的广泛使用的方法，而该技术是外部研究的金标准。此外，NTA方法可以整合荧光检测，从而识别单个样本中外泌体的亚群的能力。粒子Metrix的Zetaview Twin使用NTA技术来检测囊泡的大小和浓度，但还将检测两个不同的荧光团。这是一种能够测量从纳米尺寸到微米尺寸的囊泡的最先进的仪器，并且有助于生成高质量的高通量数据。总体而言，该工具是支持UC现有基础设施以研究外泌体运输的必要补充。特别是，Zetaview Twin对于量化体内研究的EV/外泌体是必要的。例如，药物处理是否会改变荧光标记外泌体的大小，浓度或特定亚群？在野生和驯养的鱼类中的压力或毒性暴露会导致血浆外泌体特征的变化吗？微生物释放的外泌体会影响肠道上皮细胞吗？这些只是主要申请人和共同申请者希望使用Zetaview Twin回答的一些问题。对设备的收购对于卡尔加里大学的下一阶段的电动汽车研究至关重要，并将支持高质量的HQP招募和培训，以及促进突破性研究。 **