The IMPA2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the IMPA2 gene in the human NCI-H1975 lung adenocarcinoma cell line. This product comprises a heterogeneous pool of edited cells that facilitates study of IMPA2 loss-of-function within a well-characterized cancer model. The CRISPR/Cas9-mediated gene disruption eliminates functional IMPA2, enabling researchers to interrogate its role in myo-inositol metabolism and downstream signaling pathways.
The parental NCI-H1975 line, derived from pleural effusion of a non-small cell lung cancer (NSCLC) patient, harbors EGFR L858R/T790M mutations that drive constitutive PI3K/AKT pathway activation. These cells are a standard model for EGFR-mutant lung adenocarcinoma, particularly for investigating kinase inhibitor resistance and alternative therapeutic targets. Their epithelial origin and dependency on EGFR signaling make them ideal for examining metabolic and signaling adaptations upon gene perturbation.
IMPA2 encodes an inositol monophosphatase that hydrolyzes inositol-1-phosphate to free myo-inositol, a precursor for phosphoinositide synthesis. This reaction supplies the PI metabolic pathway, generating PIP2 and PIP3, and thus sustains PI3K/AKT signaling downstream of activated EGFR. IMPA2 activity is lithium-sensitive and functions alongside IMPA1. Knockout depletes myo-inositol, limiting PIP3 production and reducing AKT phosphorylation, effectively disrupting growth factor-to-AKT signal transduction. PTEN further modulates this pathway by dephosphorylating PIP3, making IMPA2 a key metabolic regulator of EGFR-dependent PI3K/AKT output.
In NCI-H1975 cells, where EGFR mutations chronically activate PI3K/AKT, IMPA2 loss introduces a metabolic vulnerability by starving the pathway of inositol-based substrates. This model allows dissection of how myo-inositol metabolism contributes to tumor cell survival independently of direct PI3K mutations. The polyclonal nature recapitulates editing heterogeneity, providing a realistic population for studying metabolic dependencies. Additionally, because lithium inhibits IMPA2, these cells are suitable for examining lithium sensitivity and its interplay with EGFR signaling.
Applications include measuring AKT phosphorylation (p-AKT S473) by western blotting, quantifying intracellular myo-inositol levels, and assessing PI3K activity via ELISA. Cell viability (MTS/MTT) and colony formation assays probe growth dependencies, while RT-qPCR confirms gene disruption. Lithium dose-response studies can reveal compensatory roles of IMPA1. Thus, this polyclonal knockout model serves as a powerful tool for investigating inositol metabolism in EGFR-mutant lung cancer. For inquiries, contact Ascent Research.
