Protein-engineered nanostructures to illuminate protein delivery and cellular processing

细胞可以摄取具有不同形状的纳米颗粒（NP）。该项目将使用治疗性蛋白质作为构建模块来创建NP。这些蛋白质将被设计成能够自发形成柔性的细长颗粒。这些蛋白质将结合分子簇，这些分子簇将与患病细胞强烈而特异地结合。控制纳米颗粒的大小、形状和化学性质将决定它们在注射后在人体中的累积位置。如果成功，该项目可能会导致癌症和其他疾病靶向蛋白质递送的新方法。 高中生参与暑期研究机会，再加上本科生的长期研究机会，将有助于培养STEM人才。 该项目的目标是设计一种能够靶向细胞内递送蛋白质的蛋白质纳米结构平台。一个关键的挑战是，循环所需的物理和化学特性与细胞摄取所需的特性不同。纳米尺度的尺寸和“隐形”化学最适合循环，而细胞结合配体的分子尺度模式最适合细胞摄取。该项目将涉及多个步骤。模块化蛋白质工程方法将被应用于构建具有明确定义的物理特性的纳米结构，使用活性酶作为构建模块。将使用蛋白质寡聚体的非天然氨基酸（UAA）修饰来创建具有定义的配体聚集以及配体各向异性的纳米结构。这些纳米结构的物理化学性质，定义重要的细胞内交付的结果将被表征。最终，该项目将为组装成特定纳米结构的蛋白质低聚物的设计，构建和生长提供新的基本见解。期望是，这将导致新的能力设计？蛋白质材料？该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Cells can ingest nanoparticles (NPs) that have distinct shapes. This project will create NPs using therapeutic proteins as building blocks. The proteins will be designed so that they can spontaneously form flexible, elongated particles. The proteins will incorporate clusters of molecules that will bind strongly and specifically to diseased cells. Controlling the size, shape, and chemistry of the NPs will determine where they accumulate in the human body after injection. If successful, this project could lead to new methods for targeted protein delivery in cancer and other diseases. Engagement of high school students in summer research opportunities, combined with extended research opportunities for undergraduate students will help develop a STEM workforce. The project objective is to design a protein nanostructure platform capable of targeted intracellular delivery of proteins. A critical challenge is that the physical and chemical characteristics necessary for circulation are distinct from those necessary for cell uptake. Nanoscale dimensions and "stealth" chemistries are best for circulation whereas molecular-scale patterns of cell-binding ligands are best for cell uptake. The project will involve multiple steps. Modular protein engineering approaches will be applied to construct nanostructures with well-defined physical properties, using active enzymes as building blocks. Nanostructures with defined ligand clustering as well as ligand anisotropy will be created using unnatural amino acid (UAA) modification of protein oligomers. The physicochemical properties of these nanostructures that define important intracellular delivery outcomes will be characterized. Ultimately, the project will offer new fundamental insights into the design, construction, and growth of protein oligomers that assemble into specific nanoarchitectures. The expectation is that this will lead to new capabilities to design ?protein materials? that can address prohibitive barriers in systemic administration and delivery of intracellular proteins.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.