The HMGCL Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, providing a powerful loss-of-function model for studying the HMGCL gene in a human lung adenocarcinoma background. This product offers a heterogeneous pool of cells with targeted disruption of the HMGCL locus, enabling investigation of ketogenesis and leucine catabolism without the need for clonal isolation.
The parental NCI-H1975 cell line is an established human non-small cell lung cancer (NSCLC) model derived from a lung adenocarcinoma. These cells harbor activating mutations in EGFR (L858R) and T790M, making them a widely used system for studying oncogenic signaling and metabolic adaptations in cancer. Their epithelial origin and robust growth characteristics facilitate high-throughput investigations into the interplay between genetic perturbations and metabolic reprogramming.
HMGCL encodes 3-hydroxy-3-methylglutaryl-CoA lyase, a mitochondrial enzyme that catalyzes the cleavage of HMG-CoA to acetyl-CoA and acetoacetate, a critical step in ketogenesis and the final step of leucine degradation. The enzyme requires divalent cations, with magnesium and manganese serving as essential cofactors. Its activity is tightly regulated by metabolic signals: PPAR?? and glucagon promote HMGCL expression during fasting, while insulin suppresses it. Downstream, acetoacetate can be further metabolized to ketone bodies, linking HMGCL to systemic energy homeostasis. HMGCL operates in concert with HMGCS2, which generates the HMG-CoA substrate, and BDH1, which interconverts acetoacetate and 3-hydroxybutyrate, while ACAT1 mediates the reverse reaction in ketolysis. Disruption of HMGCL therefore impairs the canonical ketogenic pathway and leucine catabolism, reducing ketone body output and potentially altering acetyl-CoA pools.
In NCI-H1975 cells, knockout of HMGCL provides a unique tool to examine the metabolic vulnerabilities of lung adenocarcinoma. NSCLC cells often exhibit altered mitochondrial function and lipid metabolism to support proliferation; by eliminating HMGCL-driven ketogenesis and leucine degradation, this model reveals how cancer cells rewire metabolic fluxes when these pathways are blocked. Studies can explore compensatory mechanisms, such as increased reliance on glucose or glutamine metabolism, and assess the impact on cellular energy status and redox balance. This polyclonal population retains the EGFR signaling context of the parental line, allowing integrated analysis of oncogenic signaling and metabolic dependency.
This knockout model is ideally suited for a range of applications including ketogenesis research, cancer metabolic reprogramming, leucine metabolism studies, metabolic vulnerability profiling, and mitochondrial function studies. Representative experiments include western blotting and RT-qPCR for target validation, ketone body quantification to monitor pathway output, LC-MS metabolomics for broad metabolite profiling, mitochondrial respiration assays to assess oxidative phosphorylation, and 13C-leucine isotope tracing to map catabolic flux. By combining these approaches, researchers can elucidate the role of HMGCL in tumor metabolism and identify potential therapeutic targets. For further details, please contact Ascent Research.
