期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2015
卷号：112
期号：5
页码：1422-1427
DOI：10.1073/pnas.1424111112
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceA major challenge with in vitro investigations of the pathophysiological processes in sickle cell disease (SCD) has been the lack of a well-controlled microenvironment to mimic in vivo circulating conditions. The microfluidic platform developed here provides a quantitative assay of the kinetics of cell sickling, unsickling, and single-cell rheology. The ensuing alterations in the biorheological characteristics of sickle cells under hypoxic conditions show strong correlation with sickle hemoglobin level, hydroxyurea (HU) therapy, and cell density. These analyses provide cell-level perspectives of the clinical manifestations in SCD patients and offer unique diagnostic indicators of vasoocclusion and disease severity. These results could also provide alternative pathways to supplement current clinical practices to evaluate HU therapy. We developed a microfluidics-based model to quantify cell-level processes modulating the pathophysiology of sickle cell disease (SCD). This in vitro model enabled quantitative investigations of the kinetics of cell sickling, unsickling, and cell rheology. We created short-term and long-term hypoxic conditions to simulate normal and retarded transit scenarios in microvasculature. Using blood samples from 25 SCD patients with sickle hemoglobin (HbS) levels varying from 64 to 90.1%, we investigated how cell biophysical alterations during blood flow correlated with hematological parameters, HbS level, and hydroxyurea (HU) therapy. From these measurements, we identified two severe cases of SCD that were also independently validated as severe from a genotype-based disease severity classification. These results point to the potential of this method as a diagnostic indicator of disease severity. In addition, we investigated the role of cell density in the kinetics of cell sickling. We observed an effect of HU therapy mainly in relatively dense cell populations, and that the sickled fraction increased with cell density. These results lend support to the possibility that the microfluidic platform developed here offers a unique and quantitative approach to assess the kinetic, rheological, and hematological factors involved in vasoocclusive events associated with SCD and to develop alternative diagnostic tools for disease severity to supplement other methods. Such insights may also lead to a better understanding of the pathogenic basis and mechanism of drug response in SCD.
