Collaborative Research: Three-Dimensional Flexible Biosensor Enabling Label-Free Spatial Mapping of Intra-Organoid Functions

在过去的几十年里，生物学家和生物工程师一直试图设计“活”细胞群，这些细胞群可以在体外展示组织和器官的结构和功能。这些被称为类器官的“活”细胞团可以作为一种低成本、快速而精确的药物筛选平台，并最终取代动物模型。尽管前景看好，但仍然缺乏工具来实现对类器官生物活动的实时、非侵入性监测。该项目支持一项综合研究和教育计划，其目标是：(1)开发一种三维柔性传感器，破译人类心脏样器官的电生理作用，并用它来预测潜在的COVID-19抗病毒药物对人类心脏的影响；(2)为不同层次的学生开发与生物传感器相关的多学科教育框架。尤其是来自科学和工程领域传统上代表性不足的群体。拟议的研究将通过提供一种新的类器官传感器平台，对正在进行的COVID-19和未来不可预见的爆发确定有效的治疗方法，对美国的健康和经济产生积极和直接的影响。在过去的几十年里，人们做了大量的努力来概括器官的多细胞、解剖和功能特征，从而提供了全面的框架来模拟器官发育、体内平衡、再生和疾病。这些运动迅速形成了各种类似器官的多细胞集群，被称为类器官。然而，仍然缺乏能够无标签、实时和无创地监测类器官内功能的工具。本项目旨在建立一套材料、设计布局和组装方法，以开发一种垂直有序硅纳米针为仪器的三维柔性类器官内传感器。作为一个模型系统，该传感器将被定制为来自心血管类器官内部的心电图信号的无标记空间制图，这些器官是通过协调多能干细胞向心肌细胞和内皮细胞的空间组织共分化而设计的。类器官内活动的定量读数将有助于更好地理解心脏功能的潜在解剖-电生理-机械关系。此外，这种类器官内传感器平台将成为一种变革性的器官芯片工具，它将极大地帮助确定新开发药物的功效以及未知毒素的影响。将建立互补的实验和计算方法来分析与血管化心肌功能相关的时间序列数据。这项合作研究是建立在普渡大学和伊利诺伊大学厄巴纳-香槟分校之间长期合作关系的基础上，在过去两年中，多个pi的共同努力建立了强有力的研究联系。由于这两所大学位于中西部地区，地理位置很近，普渡大学的研究人员将能够在伊利诺伊大学厄巴纳-香槟分校的临床环境中花费大量时间，不仅可以从临床角度获得及时的反馈，还可以确保拟议任务的进展。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。

Over the last decades, biologists and bioengineers have attempted to engineer "living" cell masses that exhibit tissues and organ structure and function outside of the body. These "living" cell masses, named organoids, can serve as a low-cost, rapid, but precise drug-screening platform and, ultimately, replace animal models. Despite the promise, there remains a lack of tools to enable real-time, non-invasive monitoring of the organoids' biological activities. This project supports an integrated research and educational program with goals to (1) develop a three-dimensional flexible sensor deciphering electrophysiological actions of human heart-like organoids and use it to predict the effect of potential anti-viral drugs for COVID-19 on the human heart, and (2) develop a multidisciplinary educational framework associated with the biosensor for multiple levels of students, especially from traditionally underrepresented groups in science and engineering. The proposed research will make a positive and immediate impact on U.S. health and economy by providing a novel organoid-sensor platform useful to determine powerful therapeutics to the on-going COVID-19 and future, unforeseeable outbreak.For the last decades, extensive efforts have been made to recapitulate the multicellular, anatomical, and functional hallmarks of organs, thereby offering comprehensive frameworks to model organ development, homeostasis, regeneration, and disease. These movements have quickly engineered a variety of organ-like multicellular clusters named organoids. However, there remains a lack of tools enabling label-free, real-time, and non-invasive monitoring of intra-organoid functions. This project aims to establish a set of materials, design layouts, and assembly methods to develop a three-dimensional flexible intra-organoid sensor instrumented with vertically ordered silicon nanoneedles. As a model system, this sensor will be tailored for label-free spatial mapping of electrocardiogram signals from the inside of cardiovascular organoids that are engineered by orchestrating spatially-organized co-differentiation of pluripotent stem cells to cardiac muscle cells and endothelial cells. The quantitative readout of the intra-organoid activities will facilitate improved understanding of the underlying anatomical-electrophysiological-mechanical relationships of cardiac function. Furthermore, this intra-organoid sensor platform will become a transformative organ-on-a-chip tool that will greatly assist efforts to determine the efficacy of newly developed drugs as well as the impact of unidentified toxins. Complementary experimental and computational methods will be established for analyzing time-series data associated with vascularized cardiac muscle functions. This collaborative research has been built upon a strong research tie of the Multiple-PIs in joint efforts over the past 2 years based on a long-standing relationship between Purdue University and the University of Illinois at Urbana-Champaign. Because the universities are within close geographic proximity in the Midwest, the investigators at Purdue University will be able to spend significant amounts of time in the clinical setting at the University of Illinois at Urbana-Champaign in order to not only obtain timely feedback from the clinical perspectives but also ensure the progress of the proposed tasks.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.