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，负责将氧气从肺部输送到身体的其他部位，因此了解这些细胞是如何工作的至关重要，特别是在影响它们健康的疾病中。这项工作试图利用各种技术来解决红细胞知识中的关键空白，这些技术将探测由于红细胞健康变化而导致的内部、外部和细胞膜的变化。这将促使人们进一步了解与红细胞相关的疾病的早期变化，如疟疾、贫血和心血管疾病。除了不断进步的研究，这个职业奖项还将教育未来的劳动力和社区，他们需要做出关于新技术的明智决定。这个项目将教授学生研究概念，让他们接触到新的细胞特征和最先进的技术，并培训他们如何与公众交流他们的工作。目前的红细胞(RBC)分析通常依赖于对细胞形态或行为变化的人工观察。细胞的电测量包含有关其功能的重要信息(S)。然而，它们很难衡量。标准技术通常测量不超过几个细胞，需要熟练的技术人员来完成分析。此外，没有一种技术可以解释电池中存在的所有离子电流。尽管如此，这些测量的更广泛的影响仍然很高，特别是在药物发现和疾病进展方面。这份职业提案的目标是调查在红细胞分析和红细胞健康监测中是否存在相互关联的“细胞电子显微镜”。由于蛋白质组是细胞表达的一整套蛋白质，所以“电势”代表了细胞中存在的一整套离子电流，包括细胞表面Zeta电位、膜电位和介电(DEP)测量的膜电导、膜电容和细胞质电导。介电泳法(DEP)是一种电动技术，它依靠细胞和悬浮介质的电生理特性以及外加电场的几何形状来诱导这些细胞的运动。由于DEP不依赖于生物标记物或其他荧光标记，它已成为一种日益流行的细胞表征方法。这项工作的主要假设是，DEP参数可以强有力地描述RBC相关疾病中的一个主导因素-氧化应激引起的RBC电信号的变化，从而提供一个快速且无标记的生物标志物。如何将这些参数联系起来将增加对RBC电生理学及其在RBC相关疾病中的作用的了解。一旦理解了红细胞中的电磁场，就可以扩展到其他更复杂的细胞类型，也可以解释这些其他细胞中的行为，如影响转移性癌症细胞间相互作用的电生理变化。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.