CAREER: Investigating the Cellular Electrome as a Biomarker in Red Blood Cell Physiology and Pathology

红细胞（RBC）是人体中最丰富的细胞，负责将氧气从肺部运送到身体的其他部位，因此了解这些细胞如何工作至关重要，特别是在影响其健康的疾病中。这项工作旨在使用各种技术来解决RBC知识的关键差距，这些技术将探测由于RBC健康变化而引起的内部，外部和膜变化。这将引起对红细胞相关疾病（如疟疾、贫血和心血管疾病）早期变化的进一步了解。除了推进研究，该职业奖将教育未来的劳动力以及社区，他们需要对新技术做出明智的决定。该项目将教授学生研究概念，让他们接触新颖的细胞表征和最先进的技术，并培训他们如何与公众交流他们的工作。目前的红细胞（RBC）分析通常依赖于人工观察细胞形态或行为的变化。细胞的电测量包含有关其功能的重要信息。然而，它们很难衡量。标准技术通常测量不超过几个细胞，并且需要熟练的技术人员来完成分析。此外，没有技术能够解释细胞中存在的所有离子电流。尽管如此，这些测量的更广泛影响仍然很高，特别是在药物发现和疾病进展方面。本CAREER提案的目的是调查在RBC分析和RBC健康监测中存在的相互连接的“细胞电生理”。由于蛋白质组是由细胞表达的蛋白质的完整集合，因此“电子组”表示存在于细胞中的离子电流的完整集合，并且包括细胞表面ζ电位、膜电位和介电泳（DEP）测量的膜电导、膜电容和细胞质电导。介电电泳（DEP）是一种动电技术，其依赖于细胞和悬浮介质的电生理特性以及所施加的电场的几何形状来诱导这些细胞的运动。由于DEP不依赖于生物标志物或其他荧光标记，因此它已成为越来越受欢迎的细胞表征方法。这项工作的主要假设是，DEP参数可以鲁棒地描述由于氧化应激而引起的RBC电极的变化，氧化应激是RBC相关疾病的主导因素，提供了快速和无标记的生物标志物。这些参数之间的联系将增加对RBC电生理学及其在RBC相关疾病中的作用的理解。一旦在红细胞中理解了电分子，就可以扩展到其他更复杂的细胞类型，并且还可以解释这些其他细胞中的行为，例如影响转移性癌症中细胞与细胞相互作用的电生理学改变。该奖项反映了NSF的法定使命，并且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。

Red blood cells (RBCs) are the most abundant cell in the human body and are responsible for carrying oxygen from our lungs to the rest of our body, thus it is crucial to understand how these cells work especially in diseases that affect their well-being. This work seeks to address critical gaps in knowledge of RBCs using a variety of techniques that will probe internal, external, and membrane alterations due to changes in RBC health. This will elicit further understanding of early changes in RBC-related diseases such as malaria, anemia, and cardiovascular diseases. In addition to progressing research, this CAREER award will educate the future workforce as well as the community, who need to make informed decisions regarding new technologies. This project will teach students research concepts, expose them to novel cell characterization and state-of-the-art technologies, and train them how to communicate their work with the public. Current red blood cell (RBC) analysis often relies on manual observations to changes in cell morphology or behavior. Electrical measures of a cell contain vital information about their function(s). They are, however, difficult to measure. Standard techniques typically measure no more than a few cells and require skilled technicians to complete the analysis. Further, no technique accounts for all ionic currents present in a cell. Still, broader impact of these measurements remains high, particularly in drug discovery and disease progression. The objective of this CAREER proposal is to investigate the existence of an interconnected “cell electrome” in the analysis of RBCs and RBC health monitoring. As the proteome is the complete set of proteins expressed by a cell, the “electrome” represents the complete set of ionic currents present in a cell and includes cell surface zeta-potential, membrane potential, and Dielectrophoresis (DEP)-measured membrane conductance, membrane capacitance, and cytoplasm conductivity. Dielectrophoresis (DEP) is an electrokinetic technique that relies on the electrophysiological properties of cells and the suspending media as well as the geometry of the applied electric field to induce movement of these cells. Since DEP does not rely on biomarkers or other fluorescent labels, it has become an increasingly popular method of cell characterization. The primary hypothesis of this work is that DEP parameters can robustly describe changes in RBC electrome due to oxidative stress, a dominant factor in RBC-related diseases, providing a rapid and label-free biomarker. How these parameters are linked will increase understanding of RBC electrophysiology and its role in RBC-related diseases. The electrome, once understood in RBCs, could be expanded to other, more complicated cell types and may also explain behaviors in these other cells such as electrophysiology alterations to influence cell-to-cell interactions in metastasizing cancer.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.