The GNPDA2 Knockout NCI-H1975 Polyclonal Cells product comprises a polyclonal population of NCI-H1975 human lung adenocarcinoma epithelial cells with CRISPR/Cas9-mediated disruption of the GNPDA2 gene. This polyclonal knockout pool serves as a loss-of-function model for studying glucosamine-6-phosphate deaminase 2 in EGFR-mutant non-small cell lung cancer (NSCLC), enabling bulk-level phenotypic assessment without clonal selection bias.
The parental NCI-H1975 cell line is a well-established NSCLC model harboring EGFR L858R and T790M mutations, which together drive oncogenic signaling and confer acquired resistance to first-generation tyrosine kinase inhibitors. Its epithelial origin from a female adenocarcinoma patient makes it particularly relevant for dissecting metabolic adaptations in EGFR-driven tumors.
GNPDA2 encodes glucosamine-6-phosphate deaminase 2, catalyzing the conversion of glucosamine-6-phosphate to fructose-6-phosphate and ammonia within the hexosamine biosynthesis pathway. It functions downstream of GFPT1/2 and interacts with GNPNAT1 and NAGK to regulate UDP-N-acetylglucosamine levels for protein O-GlcNAcylation. Allosterically activated by N-acetylglucosamine 6-phosphate, GNPDA2 is transcriptionally modulated by ChREBP and SREBP-1c in response to glucose. The generated fructose-6-phosphate enters glycolysis or gluconeogenesis, connecting amino sugar metabolism to energy homeostasis. Consequently, GNPDA2 impacts OGT and OGA activities, influencing a broad range of O-GlcNAc-modified proteins.
In NCI-H1975, GNPDA2 knockout likely disturbs hexosamine flux and O-GlcNAcylation, potentially affecting cancer cell proliferation, survival, and sensitivity to EGFR-targeted agents. Given the gene??s established GWAS connections to obesity and BMI, this model enables dissection of obesity-linked metabolic pathways in a lung adenocarcinoma context. The polyclonal population allows investigation of how heterogeneous GNPDA2 loss alters bulk tumor-cell metabolism and response to therapeutic stress.
Typical applications include LC-MS-based hexosamine metabolite profiling, Seahorse extracellular flux analysis for glycolytic function, and assessment of O-GlcNAcylation by Western blot or lectin pull-down. Proliferation can be evaluated with MTT or CellTiter-Glo, apoptosis by Annexin V/PI staining, and EGFR inhibitor sensitivity via dose-response assays. Migration and invasion assays further reveal phenotypic impact, while RNA-seq provides transcriptome-wide insights into GNPDA2-dependent pathways. For more information, please contact Ascent Research.
