A Closed Loop Control System with Live Cells in the Loop

一种带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体自噬。 使用具有传感和执行功能的创新实验试验台的电池。药理结合 干预措施被广泛用于许多疾病的治疗。因此，控制理论有很大的潜力--fi 通过仔细调整多种药物的剂量传递来提高患者的幸福感 试剂(PR)。细胞内测量自噬小泡(AVs)的形成对应用的响应 将使用传统的fl荧光显微镜和Amny ImageStream Mark II成像进行各种PR的检查 流式细胞仪，在新墨西哥大学(UNM)的自噬，在fl中，疾病中的代谢 生物医学研究卓越中心(AIM中心)，全国唯一专注于自噬的Cobre，在 皮索伦蒂诺是准会员。这台最先进的仪器使用flow细胞仪和多光谱成像技术 检测细胞内AVs，提供单个细胞内AVs总数(输出)的准确计数。泛滥的flUIDIC 作为建议的一部分，将开发及时并以受控方式交付药物(投入物)的交付系统 Effort.该实验平台将为活细胞自噬的闭环控制提供概念验证的实现 实验。此外，还将开发一个完整的数学模型来描述自噬的调节。 活细胞，考虑了几种PR对动静脉动静脉形成的ff作用。我们的控制决策将在两个小时内详细说明 水平：first，从一组可用PR中选择要用于联合研究的PR；第二，剂量函数 对于组合中的每个PR，即PR作为时间的函数被提供的速率，其中时间是连续的 功能。还将开发和测试一种创新的多场景最优控制器，以优化 存在有限的实验信息和关于系统状态的不确定性。 UNM AIM中心拥有一个专家团队，他们对自噬在疾病中的作用进行了开创性的研究 在fl，他们为初级教职员工提供指导机会。作为准会员 在AIM中心，皮索伦蒂诺在专业的本地环境中处于一个独特的位置，能够获得成功。这个 这项拟议研究的长期影响将是显著的，因为它有相当大的潜力来促进科学知识的发展。fifi 在工程学和生命科学的结合点，并导致优化治疗的开发，可以积极地ff等 世界各地数百万患者的福祉。 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