Engineering glycoprotein nanocages for cellular targeting and cargo delivery

用于细胞靶向和货物递送的工程糖蛋白纳米笼

• 批准号: RGPIN-2016-06087
• 负责人: Julien, JeanPhilippe
• 金额: $ 2.77万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=629935
• 关键词: Engineering; glycoprotein; nanocages; cellular; targeting

The ability to design nanostructures has revolutionized biomedicine, bioprocesses and electronics. Although advancements in nanotechnology and protein design have been substantially widespread, certain classes of biomolecules remain understudied. Glycoproteins, in particular, require stringent production systems and some are particularly unstable – making nanotechnology development of glycoproteins challenging. It is predicted that over half of human proteins have modifications that lead to their glycosylation. Furthermore, their multimerization regulates a variety of biological processes including gene expression, activity of receptors, and cell-cell adhesion. Thus, fundamental structure/function advances related to the multimerization of glycosylated biomolecules will directly impact nanotechnology advancements. My research in protein design and glycobiology has previously shown that self-assembling protein cages can be used for the external display of a viral glycoprotein, by optimizing its recognition by antibodies. By leveraging my expertise and resources, I want to further investigate the fundamentals of protein nanoparticle assembly and use protein engineering and biophysical techniques to broaden the development of stable glycoprotein nanocages. The objective of my research program is to uncover the fundamentals of protein nanoparticle self-assembly in mammalian cells in order to engineer the transport of multimerized glycoproteins on vehicles with loaded cargo. Aim 1 is to study the self-assembly of protein cages genetically fused to two types of glycoproteins for their external display: 1) oligomeric glycoproteins, and 2) antibody glycoproteins. In Aim 2, my team will study the fundamentals of protein encapsulation in the hollow center of the glycoprotein nanocages, with the objective to discover conditions under which all components are structurally-sound and deliverable to cells. HQPs will perform extensive biophysical, structural and functional characterization of glycoprotein nanocages to ultimately provide insights into glycoprotein design and uncover the fundamentals that govern glycoprotein nanocage interactions with cell-surface receptors. The topics, strategies, infrastructure and techniques used in my research program will provide HQPs with a tailored training in nanotechnology and high-throughput protein chemistry within a world-class research environment. Glycoproteins are widely dispersed in all biological processes – therefore discoveries made in glycoprotein nanocage manipulation have the potential to be broadly applicable and impactful in all areas where these nanotechnologies are used, such as imaging, cellular biology and biomedical engineering.

设计纳米结构的能力已经彻底改变了生物医学，生物工艺和电子学。虽然纳米技术和蛋白质设计的进步已经相当普遍，但某些类别的生物分子仍然研究不足。特别是糖蛋白，需要严格的生产系统，有些是特别不稳定的，使糖蛋白的纳米技术开发具有挑战性。据预测，超过一半的人类蛋白质具有导致其糖基化的修饰。此外，它们的多聚化调节多种生物学过程，包括基因表达、受体活性和细胞-细胞粘附。因此，与糖基化生物分子的多聚化相关的基本结构/功能进展将直接影响纳米技术的进步。我在蛋白质设计和糖生物学方面的研究先前已经表明，自组装蛋白质笼可以用于病毒糖蛋白的外部展示，通过优化抗体对其的识别。通过利用我的专业知识和资源，我想进一步研究蛋白质纳米颗粒组装的基本原理，并使用蛋白质工程和生物物理技术来扩大稳定的糖蛋白纳米笼的发展。我的研究计划的目标是揭示蛋白质纳米颗粒在哺乳动物细胞中自组装的基本原理，以便在装载货物的车辆上设计多聚化糖蛋白的运输。目的1：研究蛋白笼与两种糖蛋白（1）寡聚糖蛋白和2）抗体糖蛋白的自组装。在目标2中，我的团队将研究糖蛋白纳米笼中空中心蛋白质封装的基本原理，目的是发现所有组分结构合理并可输送到细胞的条件。HQP将对糖蛋白纳米笼进行广泛的生物物理，结构和功能表征，最终提供对糖蛋白设计的见解，并揭示控制糖蛋白纳米笼与细胞表面受体相互作用的基本原理。在我的研究计划中使用的主题，策略，基础设施和技术将为HQP提供世界一流的研究环境中的纳米技术和高通量蛋白质化学的量身定制的培训。糖蛋白广泛分布在所有生物过程中-因此，在糖蛋白纳米笼操作中的发现有可能在使用这些纳米技术的所有领域（如成像，细胞生物学和生物医学工程）中广泛应用和产生影响。