A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs
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 differenc...
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| Format: | Article |
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MDPI AG
2025-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/14/8/615 |
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| author | Katherine H. Nguyen Midori L. Wasielewski Srilakshmi Yalavarthi Xianggui Qu Jason S. Knight Shuichi Takayama |
| author_facet | Katherine H. Nguyen Midori L. Wasielewski Srilakshmi Yalavarthi Xianggui Qu Jason S. Knight Shuichi Takayama |
| author_sort | Katherine H. Nguyen |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-70b328b5fa0b424b80ed8eccac1c8cea |
| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj-art-70b328b5fa0b424b80ed8eccac1c8cea2025-08-20T03:14:20ZengMDPI AGCells2073-44092025-04-0114861510.3390/cells14080615A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface WebsKatherine H. Nguyen0Midori L. Wasielewski1Srilakshmi Yalavarthi2Xianggui Qu3Jason S. Knight4Shuichi Takayama5Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USADivision of Rheumatology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USADepartment of Mathematics and Statistics, Oakland University, Rochester, MI 48309, USADivision of Rheumatology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USAWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USANeutrophil 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.https://www.mdpi.com/2073-4409/14/8/615neutrophil extracellular trapsneutrophil extracellular trap degradationassay development |
| spellingShingle | Katherine H. Nguyen Midori L. Wasielewski Srilakshmi Yalavarthi Xianggui Qu Jason S. Knight Shuichi Takayama A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs Cells neutrophil extracellular traps neutrophil extracellular trap degradation assay development |
| title | A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs |
| title_full | A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs |
| title_fullStr | A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs |
| title_full_unstemmed | A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs |
| title_short | A Mimetic Assay of Neutrophil Extracellular Trap Degradation Using YOYO-1-Stained DNA-Histone Surface Webs |
| title_sort | mimetic assay of neutrophil extracellular trap degradation using yoyo 1 stained dna histone surface webs |
| topic | neutrophil extracellular traps neutrophil extracellular trap degradation assay development |
| url | https://www.mdpi.com/2073-4409/14/8/615 |
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