The IVD Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma epithelial cell line, designed to disrupt the IVD gene. IVD encodes isovaleryl-CoA dehydrogenase, a mitochondrial flavoprotein essential for the third step in leucine catabolism. This polyclonal knockout model offers a heterogeneous genetic background, enabling robust investigation of loss-of-function effects in metabolic pathways without the clonal bias associated with single-cell derived lines. The CRISPR/Cas9-mediated gene disruption creates a valuable resource for studying branched-chain amino acid metabolism in a cancer-relevant context.
NCI-H1975 is a widely used human non-small cell lung cancer (NSCLC) cell line established from the pleural fluid of a female patient with lung adenocarcinoma. It harbors activating EGFR L858R and T790M mutations, making it a critical model for investigating EGFR-targeted therapy resistance and signal transduction. The epithelial origin and adherent growth properties of NCI-H1975 facilitate reproducible in vitro experiments, including drug response assays and metabolic analyses. This cell line is particularly relevant for studying the metabolic dependencies of lung adenocarcinomas, where altered amino acid utilization and mitochondrial function are increasingly recognized as therapeutic targets.
IVD encodes isovaleryl-CoA dehydrogenase, a mitochondrial enzyme that converts isovaleryl-CoA to 3-methylcrotonyl-CoA in leucine catabolism. Its expression is regulated by PPARA and PGC1A, and is responsive to leucine availability via mTORC1. IVD partners with ETF and ETFDH to feed electrons into complex I of the respiratory chain, linking amino acid degradation to oxidative phosphorylation. Downstream metabolites include acetoacetate and acetyl-CoA, which enter the TCA cycle. Disruption of IVD by CRISPR/Cas9 leads to accumulation of isovaleryl-CoA and organic acids, impairing mitochondrial function and inducing metabolic stress.
In the NCI-H1975 lung adenocarcinoma background, IVD knockout allows investigation of the interplay between branched-chain amino acid metabolism and oncogenic signaling. The EGFR L858R and T790M mutations in NCI-H1975 may influence metabolic dependencies; thus, disrupting leucine catabolism can reveal impacts on cancer cell viability and mitochondrial function. Accumulation of isovaleryl-CoA and altered TCA cycle flux could sensitize cells to metabolic inhibitors or modify drug responses. This model provides a tool to study metabolic reprogramming in EGFR-mutant NSCLC and identify novel vulnerabilities.
The IVD Knockout NCI-H1975 Polyclonal Cells are suitable for diverse research applications, including modeling isovaleric acidemia and other organic acidurias, studying mitochondrial dysfunction, and investigating leucine-dependent cancer metabolism. Metabolic profiling by LC-MS can quantify isovalerylglycine and TCA cycle intermediates, while Seahorse analysis assesses bioenergetic flux. Enzyme activity assays confirm IVD loss-of-function, and viability assays screen small molecules targeting metabolic vulnerabilities. These cells also support drug discovery for metabolic disorders and cancer. For further information, please contact Ascent Research.
