Department of Genomic Medicine and Infectious Diseases, J. Craig Venter Institute, La Jolla, CA, 92037, USA.
Department of Infectious Diseases, J. Craig Venter Institute, Rockville, MD, 20850, USA.
Sci Data. 2022 Nov 16;9(1):702. doi: 10.1038/s41597-022-01798-1.
Neutrophils are the most abundant type of white blood cells in humans with biological roles relevant to inflammation, and fighting off infections. Neutrophil Extracellular Traps (NETs) act as enxogenous agents controlling invasion by bacteria, viruses, fungi, metabolic, and traumatic agents. Traditionally, studies have focused on elucidating molecular and cellular pathways preceding NET formation. Here, we developed a model to decode the human genome and proteome of developted NETs. Via in vitro system to differentiate HL-60 human myeloid cell line into neutrophil extracellular trap (ecTrap) producing cells, we isolated and captured ectrap derived DNA and proteins for shotgun sequencing. The genomic sequences revealed accurate delineation of gene composition including immune response genes and mitochondrial enrichment, while providing a reference database for future interrogation. Shotgun proteomics showed global proteins in differentiated cells with specific immune pathways when compared to undifferentiated counterparts. Coupled with omics' approaches, we validated our system by functional assays and began to dissect host-microbial interactions. Our work provides a new understanding of the genomic and proteomic sequences, establishing the first human database deposition of neutrophil extracellular traps.
中性粒细胞是人类血液中最丰富的白细胞类型，其生物学功能与炎症和抗感染有关。中性粒细胞胞外诱捕网（NETs）作为一种内源性物质，可以控制细菌、病毒、真菌、代谢物和创伤性物质的入侵。传统上，研究集中在阐明 NET 形成之前的分子和细胞途径。在这里，我们开发了一种解码已开发的 NETs 的人类基因组和蛋白质组的模型。通过体外系统将 HL-60 人髓样细胞系分化为产生中性粒细胞胞外诱捕网（ecTrap）的细胞，我们分离并捕获 ecTrap 衍生的 DNA 和蛋白质进行鸟枪法测序。基因组序列揭示了基因组成的精确描绘，包括免疫反应基因和线粒体富集，同时为未来的研究提供了参考数据库。与未分化细胞相比，鸟枪法蛋白质组学显示了分化细胞中的全局蛋白质和特定的免疫途径。结合组学方法，我们通过功能测定验证了我们的系统，并开始剖析宿主-微生物相互作用。我们的工作提供了对基因组和蛋白质组序列的新认识，建立了第一个人类中性粒细胞胞外诱捕网的数据库。
