The HIBADH Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the HIBADH gene has been disrupted in the NCI-H1975 human lung adenocarcinoma epithelial cell line. This product provides a heterogeneous loss-of-function model for investigating valine catabolism and branched-chain amino acid metabolism, suitable for pooled functional studies without clonal selection bias.
NCI-H1975 is a widely used EGFR-mutant non-small cell lung cancer model, derived from a 58-year-old female non-smoker with lung adenocarcinoma. It harbors both the EGFR L858R activating mutation and the T790M gatekeeper mutation, which confer distinctive sensitivity and acquired resistance to EGFR tyrosine kinase inhibitors (TKIs), making it ideal for studying oncogenic signaling and drug resistance mechanisms.
HIBADH encodes the mitochondrial enzyme 3-hydroxyisobutyrate dehydrogenase, which catalyzes the NAD+-dependent oxidation of 3-hydroxyisobutyrate to methylmalonate semialdehyde in the valine degradation pathway. It operates downstream of BCAT2 and the BCKDH complex and upstream of ALDH6A1, propionyl-CoA carboxylase, and methylmalonyl-CoA mutase, ultimately linking valine catabolism to the TCA cycle. HIBADH is regulated by substrate availability and PPARGC1A, and it interacts with NAD+ and mitochondrial chaperones such as HSP60. Disruption of HIBADH leads to 3-hydroxyisobutyrate accumulation and reduced propionyl-CoA and TCA cycle flux.
In EGFR-mutant NCI-H1975 cells, HIBADH knockout provides a platform to examine how impaired valine catabolism influences mitochondrial metabolism and metabolic reprogramming. This model enables exploration of metabolic vulnerabilities associated with T790M-mediated drug resistance, as alterations in branched-chain amino acid metabolism may impact tumor growth and sensitivity to EGFR TKIs.
Typical applications include metabolomic profiling of valine pathway intermediates via LC-MS, mitochondrial stress testing with Seahorse analyzers, and EGFR TKI sensitivity assays. Complementary validation assays such as Western blotting and RT-qPCR for HIBADH expression, along with cell viability and proliferation studies under variable nutrient conditions, are facilitated. For further technical information, please contact Ascent Research.
