A Closed Loop Control System with Live Cells in the Loop

一种含有活细胞的闭环控制系统 这项工作首次提出开发和应用控制理论衍生的策略来调节活细胞中的自噬。 电池使用具有传感和驱动能力的创新实验测试平台。药物组合 干预措施被广泛用于治疗许多疾病。因此，控制理论具有巨大的潜力- 通过仔细调整多种药物的剂量递送， 试剂（PR）。响应于应用的自噬囊泡（AV）形成的细胞内测量 将使用传统荧光显微镜和Amnis ImageStream Mark II成像仪进行各种PR 流式细胞仪，可在新墨西哥州大学（UNM）获得 生物医学研究卓越中心（AIM中心），全国唯一一个以自噬为重点的CoBRE， PI Sorrentino是准会员。这种最先进的仪器使用流式细胞术和多光谱成像， 检测细胞内AV，提供单个细胞内总AV（输出）的准确计数。一种灌注性的麻醉剂 将开发一个及时和以受控方式提供药物（投入物）的输送系统，作为拟议的 好的。该实验平台将为活细胞自噬的闭环控制提供概念验证 实验此外，还将建立一个完整的数学模型来描述细胞自噬的调节， 活细胞，通过考虑几种PR对AV形成的影响。我们的控制决策将在两点进行解释 水平：第一，从一组可用PR中选择用于组合研究的PR;第二，剂量函数 对于组合中的每个PR，即，供应PR的速率作为时间的函数，时间是连续的 功能一个创新的多场景优化控制器也将被开发和测试，以优化控制行动， 有限的实验信息和系统状态的不确定性的存在。 UNM AIM中心拥有一个专家团队，他们对自噬的作用进行了开创性的研究， 卫生组织还为初级教员提供指导机会。作为准成员 作为AIM中心的一员，PI Sorrentino在专业的本地环境中处于独特的位置，以取得成功。的 拟议研究的长期影响将是重大的，因为它具有相当大的潜力，以促进科学知识 在工程和生命科学的联系，并导致优化疗法的发展，可以积极地治疗 全世界数百万患者的福祉。 1

A CLOSED LOOP CONTROL SYSTEM WITH LIVE CELLS IN THE LOOP This work proposes to develop and apply, for the first time, control theory-derived strategies to regulate autophagy in live cells using an innovative experimental testbed with sensing and actuating capabilities. Combinations of pharmacological interventions are used extensively for the treatment of many diseases. Therefore, there is significant potential for control theory- based treatments to enhance the well-being of patients through careful tuning of dosage delivery of multiple pharmacological reagents (PRs). Intracellular measurements of the formation of autophagy vesicles (AVs) in response to the application of various PRs will be conducted using traditional fluorescence microscopy and an Amnis ImageStream Mark II Imaging Flow Cytometer, available at the University of New Mexico (UNM) Autophagy, Inflammation, and Metabolism in Disease Center of Biomedical Research Excellence (AIM Center), the only autophagy-focused CoBRE in the nation and in which PI Sorrentino is an Associate Member. This state-of-the-art instrument uses flow cytometry and multispectral imaging to detect intracellular AVs, providing an accurate count of total AVs (the output) within individual cells. A perfusive fluidic delivery system to deliver drugs (the inputs) in time and in a controlled manner will be developed as part of the proposed effort. This experimental testbed will provide a proof-of-concept realization of closed-loop control of autophagy in live-cell experiments. In addition, an integrated mathematical model will be developed to describe the regulation of autophagy in live cells by considering the effects of several PRs on the formation of AVs. Our control decisions will be explicated at two levels: first, the choice of PRs to use in a combination study out of a set of available PRs; and second, the dosage function for each PR in the combination, i.e., the rate at which the PR is supplied as a function of time, with time being a continuous function. An innovative multi-scenario optimal controller will also be developed and tested to optimize the control actions in the presence of limited experimental information and uncertainty about the state of the system. The UNM AIM Center hosts a team of specialists who conduct groundbreaking research into the role of autophagy in inflammatory diseases and infection, and who provide mentoring opportunities for junior faculty. As an Associate Member of the AIM Center, PI Sorrentino is in a unique position within a specialized, local environment to meet with success. The long-term impact of the proposed research will be significant, as it has considerable potential to advance scientific knowledge at the nexus of engineering and life sciences, and lead to the development of optimized therapies that could positively affect the well-being of millions of patients worldwide. 1