Nguyen Katherine H, Wasielewski Midori L, Yalavarthi Srilakshmi, Qu Xianggui, Knight Jason S, Takayama Shuichi
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
The Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA.
Cells. 2025 Apr 19;14(8):615. doi: 10.3390/cells14080615.
Neutrophil extracellular traps (NETs) are not only promising biomarkers of disease, but also potential therapeutic targets. Overproduction or the improper clearance of NETs has been linked to disease severity. In vitro NET degradation assays can reveal mechanisms and degradation efficiency differences in diseased serum samples. There is a need for more convenient assays to increase the speed of NET degradation studies. This paper describes a simplified, lower variability mimetic assay with DNA-histone structures, referred to as surface webs, that performs functionally similarly to traditional NET degradation assays with increased scalability, ease of use, shorter preparation time, and lowered costs. The surface webs are created and dehydrated in a 96-well microplate that is shelf-stable, transportable, and viable for 30 days of storage at room temperature. The surface webs, compared to NETs, have similar shapes and distribution but lower intraplate variability while degrading with healthy serum and DNase I within the same timeframe. The assay can identify patient serum with reduced degradation capabilities. This assay opens new opportunities for NET-targeted drug discovery and studies on the role of NETs as modulators of disease.
中性粒细胞胞外陷阱（NETs）不仅是很有前景的疾病生物标志物，也是潜在的治疗靶点。NETs的过度产生或清除不当与疾病严重程度有关。体外NET降解试验可以揭示患病血清样本中的机制和降解效率差异。需要更便捷的试验来提高NET降解研究的速度。本文描述了一种简化的、变异性较低的模拟试验，该试验采用DNA-组蛋白结构，称为表面网，其功能与传统的NET降解试验相似，具有更高的可扩展性、易用性、更短的制备时间和更低的成本。表面网在96孔微孔板中生成并脱水，该微孔板可长期保存、便于运输，在室温下可保存30天。与NETs相比，表面网具有相似的形状和分布，但在同一时间范围内用健康血清和DNase I降解时，板内变异性更低。该试验可以识别降解能力降低的患者血清。该试验为以NETs为靶点的药物发现以及研究NETs作为疾病调节剂的作用开辟了新的机会。
