Microfluidic auto-alignment of protein patterns for dissecting multi-receptor crosstalk in platelets.
Microfluidic auto-alignment of protein patterns for dissecting multi-receptor crosstalk in platelets.
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DOI：
10.1039/c8lc00464a
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发表时间：
2018-09-26
期刊：
影响因子：
6.1
通讯作者：
Lu H
中科院分区：
文献类型：
--
作者：
Zhou F;Chen Y;Felner EI;Zhu C;Lu H
细胞粘附在许多细胞功能中起关键作用，例如止血/血栓形成过程、炎症反应和适应性免疫应答。许多细胞粘附过程涉及多个配体-受体系统之间通过细胞内信号传导的串扰。为了阐明这种串扰，需要分析多受体种类的结合和信号传导的协同或拮抗作用。这些分析的现有技术，例如，原子力显微镜（AFM）和生物膜力探针（BFP）分析要么是劳动密集型的、低通量的，要么在它们可以询问的配体类型方面受到限制。为了避免这些限制，需要一种技术来操纵配体与细胞群的相互作用并测量细胞群的功能反应。在这项工作中，我们开发了一个微流控平台，通过分离它们在空间和时间上的作用来研究多受体物种的结合和信号传导。该平台通过单个通道引导细胞，并使用顺序呈现的配体进行预处理和刺激细胞，然后报告细胞活化状态和功能后果。我们的方法在不同的空间区域精确地对多个蛋白质进行图案化，没有间隙。我们通过使用该平台在流动下血小板粘附和信号传导的背景下分析血小板受体、糖蛋白Ib和IIb-IIIa之间的串扰，证明了我们的方法的实用性。我们通过将其应用于分析全血样本并评估健康和糖尿病患者之间血小板活化的差异来展示该平台的临床实用性。
Cell adhesion plays a critical role in many cellular functions, such as the hemostatic/thrombotic process, inflammatory reactions, and adaptive immune response. Many cell adhesion processes involve crosstalk between multiple ligand–receptor systems through intracellular signaling. To elucidate such crosstalk requires analysis of the synergistic or antagonistic effects of binding and signalling of multi-receptor species. Current techniques for these analyses, e.g., atomic force microscopy (AFM) and biomembrane force probe (BFP) assays, are either labor-intensive, low-throughput, or limited in the types of ligands they can interrogate. Circumventing these limitations requires a technique for manipulating ligand interactions with and measuring the functional response of a population of cells. In this work, we have developed a microfluidic platform for studying binding and signaling of multi-receptor species by separating their actions in space and time. The platform directs cells through a single channel and uses sequentially presented ligands for pre-processing and stimulating cells, followed by reporting of cell activation states and functional consequences. Our method precisely patterns multiple proteins in different spatial regions without gaps. We demonstrate the utility of our method by using this platform to analyze the crosstalk between platelet receptors, glycoprotein Ib and IIb-IIIa, in the context of platelet adhesion and signaling under flow. We show the clinical utility of this platform by applying it to analyze whole blood samples and to assess differences in activation of platelets between healthy and diabetic patients.
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影响因子：
15.9
作者：
Yago, Tadayuki;Lou, Jizhong;Zhu, Cheng
通讯作者：
Zhu, Cheng
Von Willebrand因子-A1结构域在具有独特的力依赖性解离动力学的多个状态下结合了血小板糖蛋白IBα。
影响因子：
7.5
作者：
Ju L;Chen Y;Zhou F;Lu H;Cruz MA;Zhu C
通讯作者：
Zhu C
影响因子：
4.4
作者：
Chemnitz, JM;Parry, RV;Riley, JL
通讯作者：
Riley, JL
影响因子：
64.8
作者：
KIM, E;XIA, YN;WHITESIDES, GM
通讯作者：
WHITESIDES, GM
影响因子：
4.8
作者：
Bunch, Thomas A.
通讯作者：
Bunch, Thomas A.