The HADHB Knockout NCI-H1975 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted HADHB in the NCI-H1975 lung adenocarcinoma background. This polyclonal format captures diverse gene-editing outcomes, providing a robust model for loss-of-function studies without requiring clonal isolation. The cells are designed for researchers investigating mitochondrial fatty acid oxidation and its roles in cancer metabolism.
NCI-H1975 is an epithelial human non-small cell lung cancer (NSCLC) cell line harboring activating EGFR L858R and resistance T790M mutations. This genotype is clinically relevant for studying oncogenic signaling and metabolic adaptations associated with tyrosine kinase inhibitor therapy. The EGFR mutational status makes NCI-H1975 a valuable host for exploring how mitochondrial metabolism intersects with growth factor signaling in lung adenocarcinoma.
HADHB encodes the beta subunit of the mitochondrial trifunctional protein (MTP), which partners with HADHA to catalyze the thiolytic cleavage of long-chain 3-ketoacyl-CoA into acetyl-CoA and shortened acyl-CoA, the final step of fatty acid ??-oxidation. HADHB expression is transcriptionally regulated by PPAR?? and PPAR??, coactivated by PGC-1?? (PPARGC1A), and responsive to energy sensors including AMPK, SIRT1, and FOXO1. Knockout impairs long-chain fatty acid oxidation, leading to accumulation of long-chain acylcarnitines and reduced synthesis of acetyl-CoA, TCA cycle intermediates, ketone bodies (acetoacetate, ??-hydroxybutyrate), and ATP, while increasing reactive oxygen species. This disruption profoundly affects mitochondrial energy metabolism and lipid homeostasis.
In the EGFR-mutant NCI-H1975 context, HADHB loss recapitulates key features of mitochondrial trifunctional protein deficiency, linking impaired fatty acid oxidation to cancer cell metabolism. NSCLC cells often rely on metabolic reprogramming, and this knockout forces a shift away from long-chain lipid utilization, creating potential metabolic vulnerabilities. The model is suited for studying crosstalk between EGFR signaling and lipid metabolism under metabolic stress or therapeutic pressure from tyrosine kinase inhibitors.
Applications include cancer metabolism studies, investigation of fatty acid oxidation in NSCLC, and metabolic reprogramming in EGFR-mutant lung adenocarcinoma. Suitable assays encompass 14C-palmitate oxidation to measure ??-oxidation flux, acylcarnitine profiling by LC-MS, Seahorse mitochondrial respiration analysis, ATP measurement, ROS detection, and lipid accumulation staining. Confirmatory methods include Western blotting for HADHB and RT-qPCR. The cells enable drug sensitivity assays targeting ??-oxidation and metabolic adaptation to tyrosine kinase inhibitors. For further details, contact Ascent Research.
