The GNPNAT1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. These cells harbor targeted disruption of the GNPNAT1 gene, which encodes glucosamine-6-phosphate N-acetyltransferase 1, a key enzyme in the hexosamine biosynthesis pathway. The polyclonal format provides a heterogeneous pool of edited alleles, enabling studies of gene function without clonal selection artifacts. This product is designed for advanced biomedical research into glycosylation-dependent signaling in non-small cell lung cancer (NSCLC).
The NCI-H1975 parental cell line is a widely used model of NSCLC, originally established from the pleural effusion of a female patient with lung adenocarcinoma. These cells harbor activating EGFR mutations L858R and T790M, the latter conferring resistance to first-generation tyrosine kinase inhibitors (TKIs). Consequently, NCI-H1975 is particularly relevant for investigating TKI resistance and EGFR-driven oncogenic signaling in lung cancer.
GNPNAT1 catalyzes the acetylation of glucosamine-6-phosphate to N-acetylglucosamine-6-phosphate, a rate-limiting step in the hexosamine biosynthesis pathway that yields the essential donor substrate UDP-GlcNAc. UDP-GlcNAc is critical for N-linked and O-linked glycosylation as well as O-GlcNAcylation, a dynamic post-translational modification regulating protein function. GNPNAT1 activity is influenced by upstream regulators such as glucose and glutamine availability, EGFR signaling, and the transcription factor Sp1. Downstream, GNPNAT1 controls the UDP-GlcNAc pool, which drives O-GlcNAcylation of key oncoproteins including c-Myc, p53, and AKT, and modulates signaling pathways such as NF-??B and FOXO. GNPNAT1 functions in concert with interacting factors like GFPT1, NAGK, OGT, and OGA, and forms a homodimer. Disruption of GNPNAT1 therefore reduces UDP-GlcNAc levels, impairing both O-GlcNAcylation and N-glycan synthesis.
In the NCI-H1975 EGFR-mutant lung cancer context, GNPNAT1 knockout disrupts the hexosamine biosynthesis pathway and likely attenuates O-GlcNAcylation of EGFR downstream effectors, potentially altering cellular proliferation, survival, and drug sensitivity. This model offers a unique tool to dissect how metabolic flux through the hexosamine pathway intersects with mutant EGFR signaling, and to explore the contribution of protein glycosylation to TKI resistance and tumor metabolism in NSCLC.
This polyclonal knockout pool is suitable for a range of investigations, including metabolic vulnerability analysis where researchers can assess the impact of reduced UDP-GlcNAc on cell growth and apoptosis using proliferation and apoptosis assays. It enables the study of O-GlcNAcylation dynamics in NSCLC through Western blotting for O-GlcNAc and GNPNAT1, RT-qPCR, and LC-MS quantification of UDP-GlcNAc. The model can be applied to screen glycometabolic therapeutics, evaluate drug sensitivity to EGFR inhibitors, and examine migration/invasion phenotypes altered by glycosylation changes. Additionally, immunofluorescence and lectin blotting can characterize alterations in glycosylation patterns. For further details or to inquire about this product, please contact Ascent Research.
