The GRHPR Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from human A-549 lung adenocarcinoma cells, featuring targeted disruption of the GRHPR gene. This heterogeneous knockout model enables loss-of-function studies without clonal isolation, providing a robust system for investigating glyoxylate metabolism and oxalate-induced cytotoxicity in a cancer cell context.
A-549 cells, originally isolated from a 58-year-old Caucasian male with lung adenocarcinoma, serve as a well-characterized model of alveolar Type II epithelium. Their adherent morphology and stable growth facilitate genetic manipulation, while their cancer origin allows exploration of metabolic alterations relevant to tumor biology and metabolic disorders.
GRHPR encodes glyoxylate reductase/hydroxypyruvate reductase, an NADPH-dependent enzyme that reduces glyoxylate to glycolate and hydroxypyruvate to D-glycerate. In knockout cells, loss of this activity leads to glyoxylate accumulation, which is diverted to oxalate through alternative pathways, causing cellular toxicity. The accompanying reduction in D-glycerate impairs serine biosynthesis, affecting one-carbon metabolism. Key pathway components include the substrates glyoxylate and hydroxypyruvate, the cofactor NADPH, and related enzymes HAO1 and AGXT, which collectively regulate oxalate and glycine/serine metabolism.
In the A-549 background, GRHPR knockout perturbs metabolic homeostasis, potentially sensitizing cells to oxidative and nutritional stress. This model recapitulates biochemical features of primary hyperoxaluria type 2 (PH2), a disease marked by excessive oxalate production and kidney injury. By combining the lung adenocarcinoma phenotype with GRHPR deficiency, researchers can examine cell-autonomous effects on proliferation, detoxification capacity, and metabolic flux, offering insights into cancer metabolism and PH2 pathogenesis.
Research applications include modeling PH2-related nephrotoxicity via oxalate stress assays, metabolomic profiling by LC-MS for glyoxylate and oxalate quantification, and functional validation by Western blotting and RT-qPCR. The polyclonal pool is suited for drug screening to identify modulators of oxalate synthesis or serine pathway rescue. Proliferation and viability studies under varied nutrient conditions can reveal metabolic vulnerabilities. For more information or to discuss custom cell-based services, please contact Ascent Research.
