Validation of Imaging and Electrical Impedance-Based Microfluidic Assays for Cell Sickling

β-珠蛋白基因的单一突变会导致异常镰状珠蛋白(HBs)的产生， 导致镰状细胞病(SCD)。HBS在低氧气张力下聚合成刚性纤维， 导致扭曲、僵硬的红细胞(称为细胞镰状)，并导致血液流变学异常， 血管闭塞危象、溶血性贫血等。细胞镰状细胞可作为诊断生物标志物 和一个监测生物标记物来评估化疗和基因治疗的疗效 直接或间接地针对哈佛商学院及其聚合。我们最近开发了一种 基于微流控的低氧芯片处理小体积血液样本(~少μL)和 通过使红细胞处于低氧压下，在体外快速(~几分钟)诱导细胞病态。 使用细胞成像或基于阻抗的方法，无需标记即可实现对细胞病态的评估 方法：研究方法。在概念化项目中，我们将使用SPILED来研究化验灵敏度 去白细胞的纯合子镰状细胞样本。在下一阶段，我们将在体外验证这两种方法 使用纯合子和杂合子样品进行检测，建立相关的操作标准， 并自动执行数据处理和分析。该项目将导致有效的体外检测 快速、无标记测量细胞镰状细胞以评估患者对基因治疗的反应 一般针对HBS聚合的治疗和其他治疗。

A single mutation in the β-globin gene results in production of abnormal sickle globin (HbS), leading to sickle cell disease (SCD). HbS polymerizes into rigid fibers under low O2 tension, causing distorted, rigid red cells (known as cell sickling), and leading to abnormal blood rheology, vasooccclusive crisis, and hemolytic anemia etc. Cell sickling can serve as a diagnostic biomarker of SCD and a monitoring biomarker to assess the efficacy of chemotherapy and gene therapies that target HbS and its polymerization, directly and indirectly. We have recently developed a microfluidics-based hypoxia chip to process small volume of blood specimens (~ few μL) and induce cell sickling in vitro rapidly (~ few min) by subjecting red blood cells to low O2 tension. Assessment of cell sickling is achieved label-free, using cell-imaging or electrical impedance-based methods. In the conceptualization project, we will study assay sensitivity using spiked leukoreduced homozygous sickle cell samples. In the next stage, we will validate both in vitro assays using homozygous and heterozygous samples, establish relevant operating standards, and automate data processing and analysis. The project will result in validated in vitro assays for rapid, label-free measurement of cell sickling to assess patient response to gene-therapy treatments and other treatments targeting HbS polymerization in general.