Collaborative Research: CPS: Medium: CyberOrganoids: Microrobotics-enabled differentiation control loops for cyber physical organoid formation

该项目旨在创建一个网络物理系统，用于实时远程控制细胞过程，并利用合成生物学和微型机器人的生物医学潜力来创建胰腺组织。目前仅在美国就有114,000人在等待挽救生命的器官移植，直接生产患者相容器官的能力，避免了对动物和临床研究的需要，可以彻底改变个性化医疗。人体组织，如肝脏、肾脏和胰岛，包括以复杂模式排列的细胞，跨越二维和三维结构。然而，基于支架和微凝胶的组织工程方法以及3D生物打印通常无法创建这些复杂的3D结构。在这个项目中，该团队专注于胰腺，它具有独特的解剖结构，由称为胰岛的圆形细胞团的规则排列组成。提出的研究旨在通过开发一种网络物理过程来克服在体外重建这些空间模式的障碍，通过该过程，微机器人群将在3D中被引导来调节基因工程干细胞的分化并驱动这些细胞形成所需的胰腺组织。这项工作的广泛影响是重要的，因为它是合成新的或修复生病的人体器官的关键的第一步，为计算机控制的微型机器人和基因编程的干细胞之间的交互行为提供了条件。制造活组织是革命性的，因为它可以作为临床前和临床试验之间的桥梁，以确保更好的药物测试模型和开发更个性化的精准医疗。特别是对于胰腺成分，产生符合制药标准的人类类器官是一个特殊的挑战，目前的方法是费力的，耗时的，昂贵的，不可复制的，这导致工业界回避这种器官。作为该项目的研究组成部分的补充，教育和推广活动解决了在解决问题的研究职业中增加代表性不足的少数民族（即妇女和服务不足的人群）的需求，例如K-12中的工程学。与同质细胞培养相比，人类诱导多能干细胞（hiPSC）衍生的3D类器官为开发新疗法提供了更复杂和全面的模型，而不是补充动物试验。目前迫切需要开发可靠的、可扩展的类器官生产技术。该项目旨在克服这一巨大挑战，通过一种独特而新颖的网络物理系统，其中微型机器人以闭环方式增强生物系统，以实现细胞特异性功能和用户定义的时间-指导细胞命运导致类器官的形成。受“人在环路”方法的启发，我们提出了一种“μ机器人在环路”方法，在这种方法中，细胞之间的物理信号被微机器人控制的输入所取代，以提供时空精度和反馈控制来指导细胞行为。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。

This project aims to create a cyber physical system for remotely controlling cellular processes in real time and leverage the biomedical potential of synthetic biology and microrobotics to create pancreatic tissue. With 114,000 people currently on the waitlist for a lifesaving organ transplant in the United States alone, the ability to directly produce patient-compatible organs, obviating the need for animal and clinical studies can revolutionize personalized medicine. Tissues in the human body such as liver, kidney, and pancreatic islets comprise cells arranged in complex patterns spanning both 2D and 3D structures. However, scaffold- and microgel-based tissue engineering approaches along with 3D bioprinting are often unable to create these complex 3D structures. In this project, the team focuses on the pancreas, which has a unique anatomical structure composed of the regular arrangement of circular cell clusters called islets. The proposed research aims at overcoming the hurdle of recreating these spatial patterns in vitro by developing a cyber physical process by which swarms of microrobots will be steered in 3D to regulate the differentiation of genetically engineered stem cells and drive these cells into forming desired pancreatic tissue. The broader impacts of this line of work are significant because it is a key first step in the synthesis of new, or the repair of ailing, human organs, providing for interactive behavior between computer controlled microrobots and genetically programmed stem cells. Manufacturing living tissue is revolutionary as it could act as a bridge between preclinical and clinical trials, to ensure better drug testing models and develop more personalized precision medicine. For pancreatic components, in particular, generating human organoids compliant with pharmaceutical standards is an exceptional challenge, and current methods are laborious, time-consuming, expensive, and irreproducible, which has caused industry to shy away from this organ. The education and outreach activities that complement the research component of this project address the need to increase underrepresented minorities (that is, women and under-served populations) in problem-solving research careers, like Engineering in K-12. Compared to homogeneous cell cultures, human induced pluripotent stem cell (hiPSC) derived 3D organoids offer more complex and comprehensive models for developing new therapies instead of, or to complement, animal testing. There is a critical need to develop techniques for the reliable and scalable production of organoids. This project aims to overcome this great challenge, via a unique and novel cyber-physical system in which microrobots augment the biological system, in a closed-loop approach, to enable cell-specific functionality and user-defined timing – to direct cellular fate leading to the formation of organoids. Inspired by “human-in-the-loop” approaches for engineering systems that must interact with complex, living individuals, we propose a “μrobot-in-the-loop” approach in which physical signaling among cells is substituted with microrobot-controlled inputs to afford spatiotemporal precision and feedback control in directing cell behavior.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.