CAREER: A systems approach to create multiplexed microfluidics to study human immune cell dynamics

在活人身上直接研究免疫细胞的功能是很困难的。作为一种解决方案，工程师们使用小型微流体装置，也称为微生理系统，来模拟人类的健康和疾病状况。这个CAREER项目将导致多种组织微环境中人类炎症的验证模型的发展。通过模拟组织和器官的复杂3D微环境，创建的模型将使研究人员能够研究免疫细胞，信号分子和病原体（引起疾病的微生物）之间的动态相互作用。作为微流控模型的补充，数学模型提供了一个定量的框架来理解炎症的潜在机制和动力学。通过使用数学方程和计算模拟，研究人员可以设计更好的基于人类的体外（实验室）模型，并分析炎症中涉及的细胞和分子相互作用。这些模型有助于预测和优化治疗结果，揭示慢性炎症的复杂性，并确定潜在的干预措施。研究者将通过互动免疫学虚拟现实游戏将研究与教学结合起来，教授免疫系统和疫苗的工作原理。这个虚拟现实学习模块将在电气和电子工程师协会（IEEE）会议上展示，并通过与达拉斯和理查森独立学区的K-12合作伙伴关系向当地社区传播。研究者提出了一个全面的策略来招募和留住代表性不足的学生，并在达拉斯社区担任过几个领导角色，包括担任科学和工程青年女性调查员（YWISEI）计划和乔治·a·杰弗里纳米探险家计划在高中水平的导师。研究者实验室在未来十年的愿景是将数学和多路微流控实验方法结合成一个统一而强大的工具，以破译控制免疫细胞分化和激活的紧急特性，以响应外部微环境。了解不同微环境下免疫细胞功能的基本差异有望揭示终末器官衰竭的机制。为了实现这一愿景，本CAREER项目的目标是采用系统免疫学方法，将实验数据与计算建模相结合，通过追求以下目标来揭示免疫细胞反应的动力学：(1)创建一种多路微环境芯片，该芯片由定量框架（确定性和随机时空模型和数据分析方法）提供信息，用于特定类器官微环境（肺、肾、脑）中的免疫细胞迁移和中性粒细胞胞外陷阱释放（NETosis）；(2)通过将体外免疫细胞表型（迁移和NETosis）与儿科患者心脏搭桥手术前后的临床结果（终末器官衰竭）相关联的延时视频，验证芯片上多器官微环境和数学模型；(3)利用脂质介质和新型趋化微粒在芯片上调节免疫功能。本项目开发的工程工具不仅可以应用于临床，还可以应用于研究免疫细胞在炎症疾病、传染病和再生治疗中的基本机制和作用。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。

It is difficult to study the function of immune cells directly in a living person. As a solution, engineers use small microfluidic devices, also known as microphysiological systems, to simulate the conditions of health and disease in humans. This CAREER project will lead to the development of validated models of human inflammation in multiple tissue microenvironments. By mimicking the complex 3D microenvironments of tissues and organs, the models created will allow researchers to investigate the dynamic interactions between immune cells, signaling molecules, and pathogens (micoorganisms that cause disease). Complementary to microfluidic models, mathematical modeling offers a quantitative framework to understand the underlying mechanisms and dynamics of inflammation. By employing mathematical equations and computational simulations, researchers can design better human-based in vitro (in the lab) models and analyze cellular and molecular interactions involved in inflammation. These models aid in predicting and optimizing treatment outcomes, unraveling the complexity of chronic inflammation, and identifying potential interventions. The Investigator will integrate research with teaching through an interactive immunology virtual reality game that teaches how the immune system and vaccines work. This virtual reality learning module will be presented at the Institute of Electrical and Electronics Engineers (IEEE) conference and disseminated to the local community in a K-12 partnership with Dallas and Richardson Independent School Districts. The Investigator proposes a comprehensive strategy to recruit and retain underrepresented students and has served in several leadership roles in the Dallas community, including serving as a mentor for the Young Women In Science and Engineering Investigators (YWISEI) program and the George A. Jeffrey NanoExplorers Program at the high school level. The vision of the Investigator’s Lab in the next decade is to combine mathematical and multiplexed microfluidic experimental approaches into a unified and powerful tool to decipher the emergent properties governing immune cell differentiation and activation in response to the external microenvironment. Understanding the basic differences in immune cell function in different microenvironments is expected to shine light onto the mechanisms of end-organ-failure. Towards this vision, the goal of this CAREER project is to employ a systems immunology approach, integrating experimental data with computational modeling to unravel the dynamics of immune cell responses by pursuing the following objectives: (1) Creating a multiplexed-microenvironment chip that is informed by a quantitative framework (deterministic and stochastic spatiotemporal models and data analytics methods) for immune cell migration and neutrophil extracellular trap release (NETosis) in specific organoid microenvironments (lung, kidney, brain); (2) Validating the multiplexed organ microenvironments-on-a-chip and mathematical models by correlating time-lapse movies of in vitro immune cells phenotypes (migration and NETosis) with clinical outcomes (end-organ-failure) in pediatric patients before and after cardiac bypass surgery; and (3) Modulating the immune function on-chip with lipid mediators and novel chemotactic microparticles. The engineering tools developed in this project, can be applied not only clinically, but to study basic mechanisms and roles of immune cells in inflammatory disorders, infectious disease, and regenerative therapy.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.