Developing Functional Ferritin Nanocages for Blood Brain Barrier Traversing and Cellular Delivery

血脑屏障位于中枢神经系统和外周血循环之间，可以防止潜在毒素进入大脑，但也会抑制药物的成功输送。这妨碍了对致命的脑相关疾病如阿尔茨海默病、帕金森病和脑癌的有效成像、诊断和治疗。因此，对于能够解决这些问题并有效治疗脑部疾病的输送系统有着巨大的需求。铁蛋白等天然物质已被证明可以自组装成纳米笼并穿过血脑屏障。然而，对这些纳米笼如何穿过屏障进入大脑以及它们的结构-功能关系的完整理解仍然是难以捉摸的。该研究项目将结合生物工程和化学方法来了解这些纳米结构如何与血脑屏障相互作用，并合理设计具有穿越血脑屏障和实现细胞递送能力的纳米笼。该项目将提供一个强大的纳米粒子工具来研究纳米材料与生物屏障的相互作用，并将推动下一代大脑药物输送和成像平台和技术的发展。该项目汇集了几个关键技术领域，包括纳米材料、化学工程、细胞生物学和生物医学工程，为不同群体的研究生和本科生提供跨学科培训，并让当地高中的学生接触到前沿研究。该研究项目的中心目标是利用铁蛋白作为模型蛋白，全面了解基于蛋白质的纳米笼/血脑屏障相互作用，并随后利用这些知识开发一种新型蛋白质纳米笼家族，作为大脑靶向，具有额外功能的模块化交付平台。核心假设是，通过合理的设计，工程铁蛋白可以保持其组装成纳米容器的能力，穿过血脑屏障的生物界面，并且还具有靶向递送，装载治疗药物和/或显像剂的新能力。该研究项目旨在：1)设计并遗传引入铁蛋白靶向结构域，将铁蛋白组装成铁蛋白纳米笼；2)利用细胞模型研究纳米笼的血脑屏障穿越和细胞靶向能力；3)研究纳米笼结构与生物功能的关系。该项目的范围包括探测蛋白质纳米笼与生物系统的相互作用，以阐明基因导向的纳米级模块化设计，并为许多脑部疾病的潜在治疗提供诊断数据。该项目的最终目标是开发一种基因设计和化学工程的纳米级输送系统，该系统可以跨越难以穿透的血脑屏障输送治疗和诊断药物。总的来说，这些实验将探索纳米尺度的转运机制，并探索工程纳米笼与生物系统之间的相互作用，从而为设计和生产基于蛋白质的治疗系统的新范例奠定基础。该提案的教育部分将涉及来自代表性不足群体的青年，目标是吸引、激励和培训下一代STEM专业人员。该奖项反映了美国国家科学基金会的法定使命，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。

The blood-brain barrier stands between the central nervous system and peripheral blood circulation, preventing entry of potential toxins into the brain but also inhibiting the successful delivery of drugs. This prevents the effective imaging, diagnosis and treatment of deadly brain-related diseases such as Alzheimer’s, Parkinson’s disease and brain cancer. As such, there is a tremendous demand for delivery systems which can solve these problems and effectively treat brain disease. Natural materials such as ferritin proteins have been shown to self-assemble into nanocages and cross the blood-brain barrier. However, a complete understanding of how these nanocages move across the barrier into the brain as well as their structure-function relationships has remained elusive. This research project will combine bioengineering and chemical approaches to understand how these nanostructures interact with the blood-brain barrier and to rationally design nanocages which have the capability to cross blood-brain barriers and to achieve cellular delivery. The project will provide a robust nanoparticle tool to study nanomaterial-biological barrier interactions and will advance the science for the development of next generation brain drug delivery and imaging platforms and techniques. This project brings together several key technology areas, including nanomaterials, chemical engineering, cell biology and biomedical engineering, providing interdisciplinary training for a diverse group of graduate and undergraduate students as well as exposing students from local high schools to cutting-edge research. The central goal of the research project is to achieve a comprehensive understanding of protein based nanocage/blood-brain barrier interactions using ferritin as a model protein and subsequently to use this knowledge to develop a novel family of protein nanocages as brain targeted, modular delivery platforms with additional functionalities available on demand. The central hypothesis is that with rational design, the engineered ferritin protein can maintain its ability to assemble into nanocages, cross the biological interface of the blood-brain barrier, and also be conferred with new abilities for targeted delivery, loading of therapeutics and/or imaging agents. The research project seeks to: 1) design and genetically introduce targeting domains to ferritin proteins, assemble the ferritin proteins into ferritin nanocages, 2) investigate the nanocages’ blood-brain barrier traversing and cellular targeting capabilities using cellular models; and 3) study the nanocage structure-biological function relationship. The scope of the project includes probing protein nanocage interactions with biological systems to elucidate the genetically-directed nanoscale modular design and providing theragnostic data on the potential treatment of many brain diseases. The ultimate goal of the project is to develop a genetically designed and chemically engineered nanoscale delivery system which can deliver therapeutic and diagnostic agents across the impenetrable blood-brain barrier. Overall, the experiments will probe nanoscale transport mechanisms and explore the interactions between engineered nanocages and biological systems, therefore laying the foundation for design and production of a new paradigm of protein-based theranostic systems. The educational components of this proposal will involve youth from underrepresented groups with a goal to attract, inspire and train the next generation of STEM professionals.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.