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。该项目将使用治疗性蛋白质作为构建块来创建NPs。这些蛋白质的设计将使它们能够自发地形成灵活、细长的颗粒。这些蛋白质将结合成一簇簇的分子，这些分子将强烈地结合到疾病细胞上，并专门与疾病细胞结合。控制NPs的大小、形状和化学成分将决定它们在注射后在人体内积累的位置。如果成功，该项目可能会带来针对癌症和其他疾病的靶向蛋白质输送的新方法。让高中生参与暑期研究机会，加上为本科生提供更多的研究机会，将有助于培养STEM劳动力。该项目的目标是设计一种能够在细胞内靶向输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物理和化学特性不同。纳米尺度和“隐形”化学最适合循环，而分子尺度的细胞结合配体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物理性质的纳米结构，使用活性酶作为构建块。利用蛋白质低聚体的非天然氨基酸(UAA)修饰将创建具有明确的配体簇集以及配体各向异性的纳米结构。将描述这些纳米结构的物理化学性质，这些结构定义了重要的细胞内给药结果。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、建造和生长提供新的基本见解。人们的期望是，这将带来新的设计能力？蛋白质材料？这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.