Engineering glycoprotein nanocages for cellular targeting and cargo delivery

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广泛传播，但某些种类的生物分子仍未得到充分研究。糖蛋白尤其需要严格的生产系统，而且有些糖蛋白特别不稳定，这使得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有导致其糖基化的修饰。此外，它们的多聚调节多种生物过程，包括基因表达、受体活性和细胞-细胞粘附。因此，与糖基化生物分子多聚相关的基本结构/功能进步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已经表明，通过优化抗体对病毒糖蛋白的识别，自组装蛋白笼可以用于病毒糖蛋白的外部展示。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础，并利用蛋白质工程和生物物理技术来扩大稳定糖蛋白纳米笼的开发。我的研究项目的目标是揭示哺乳动物细胞中蛋白质纳米颗粒自组装的基本原理，以便在装载货物的车辆上设计多聚糖蛋白的运输。目的1是研究两种糖蛋白基因融合的蛋白笼的自组装：1)寡聚糖蛋白，2)抗体糖蛋白。在Aim 2中，我的团队将研究糖蛋白纳米笼中空中心的蛋白质封装基础，目的是发现所有成分结构健全并可传递给细胞的条件。HQPs将对糖蛋白纳米笼进行广泛的生物物理、结构和功能表征，最终为糖蛋白设计提供见解，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。在我的研究项目中使用的主题、策略、基础设施和技术将在世界级的研究环境中为HQPs提供纳米技术和高通量蛋白质化学方面的量身定制的培训。糖蛋白广泛分布在所有生物过程中，因此糖蛋白纳米笼操作的发现有可能在使用这些纳米技术的所有领域广泛应用和影响，例如成像，细胞生物学和生物医学工程。

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.