LMNA Reduced Acquired Resistance to Erlotinib in NSCLC by Reversing the Epithelial–Mesenchymal Transition via the FGFR/MAPK/c-fos Signaling Pathway

Abstract

For patients exhibiting non-small-cell lung cancer (NSCLC) with activating epidermal growth factor receptor (EGFR) mutations, epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are a first-line treatment. However, most patients who initially responded to EGFR-TKIs eventually developed acquired resistance, limiting the effectiveness of therapy. It has long been known that epithelial–mesenchymal transition (EMT) leads to acquired resistance to EGFR-TKIs in NSCLC. However, the mechanisms underlying the resistance dependent on EMT are unknown. This research aimed to reveal the effects of LMNA in the regulation of acquired resistance to erlotinib by EMT in NSCLC. The acquired erlotinib-resistant cells (HCC827/ER) were induced by gradual increase of concentrations of erlotinib in erlotinib-sensitive HCC827 cells. RNA sequencing and bioinformatics analysis were performed to uncover the involvement of LMNA in the EMT process that induced acquired resistance to erlotinib. The effect of LMNA on cell proliferation and migration was measured by clone-formation, wound-healing, and transwell assays, respectively. The EMT-related protein, nuclear shape and volume, and cytoskeleton changes were examined by immunofluorescence. Western blot was used to identify the underlying molecular mechanism of LMNA regulation of EMT. HCC827/ER cells with acquired resistance to erlotinib underwent EMT and exhibited lower LMNA expression compared to parental sensitive cells. LMNA negatively regulated the expression of EMT markers; HCC827/ER cells showed a significant up-regulation of mesenchymal markers, such as CDH2, SNAI2, VIM, ZEB1, and TWIST1. The overexpression of LMNA in HCC827/ER cells significantly inhibited EMT and cell proliferation, and this inhibitory effect of LMNA was enhanced in the presence of 2.5 μM erlotinib. Furthermore, a decrease in LMNA expression resulted in a higher nuclear deformability and cytoskeletal changes. In HCC827/ER cells, AKT, FGFR, ERK1/2, and c-fos phosphorylation levels were higher than those in HCC827 cells; Furthermore, overexpression of LMNA in HCC827/ER cells reduced the phosphorylation of AKT, ERK1/2, c-fos, and FGFR. In conclusion, our findings first demonstrated that downregulation of LMNA promotes acquired EGFR-TKI resistance in NSCLC with EGFR mutations by EMT. LMNA inhibits cell proliferation and migration of erlotinib-resistant cells via inhibition of the FGFR/MAPK/c-fos signaling pathway. These findings indicated LMNA as a driver of acquired resistance to erlotinib and provided important information about the development of resistance to erlotinib treatment in NSCLC patients with EGFR mutations.

FGFR families: biological functions and therapeutic interventions in tumors

“…In tumor cells, FGFR inhibits the JAK/STAT signaling pathway activated by T cell‐produced interferon gamma and then reduces the expression of its target genes, namely, beta‐2 microglobulin (B2M), C‐X‐C motif chemokine ligand 10 (CXCL10), and programmed death‐ligand 1 (PD‐L1), thereby mediating immune escape 60 . In non‐small cell lung cancer (NSCLC), FGFR activates the MAPK/c‐Fos pathway to promote tumor cell proliferation, migration, and resistance to erlotinib 61 . Similarly, FGFR promotes cholangiocarcinoma cell progression and resistance to gemcitabine through upregulation of AKT/mammalian target of rapamycin (mTOR) and STAT3 signaling 62 .…”

“… 60 In non‐small cell lung cancer (NSCLC), FGFR activates the MAPK/c‐Fos pathway to promote tumor cell proliferation, migration, and resistance to erlotinib. 61 Similarly, FGFR promotes cholangiocarcinoma cell progression and resistance to gemcitabine through upregulation of AKT/mammalian target of rapamycin (mTOR) and STAT3 signaling. 62 The AKT/SRY‐box 2 (SOX2) axis, mediated by FGFR2, also controls the stemness of pancreatic cancer.…”

Targeting FGFRs by pemigatinib induces G1 phase cell cycle arrest, cellular stress and upregulation of tumor suppressor microRNAs

“…Unfortunately, the onset of acquired resistance is a common event and dramatically Pace et al Journal of Translational Medicine (2023) 21:626 hampers therapeutic efficacy [63]. FGFR signaling pathways have been described as molecular mechanisms for acquired resistance to TKIs targeting other tyrosine kinase receptors, such as EGFR, c-Met, ALK and CDK4/6 [64][65][66]. Although functional studies are needed, the evidence that pemigatinib treatment can modulate other TKI targets reinforces the biological rationale for combinatorial strategies to overcome TKI resistance and improve clinical benefit.…”

Shake It Up Baby Now: The Changing Focus on TWIST1 and Epithelial to Mesenchymal Transition in Cancer and Other Diseases

“…Several studies have examined resistance to this approach in NSCLC. Hu et al compared NSCLC cells sensitive and resistant to the EGFR inhibitor erlotinib and found that resistant cells exhibited increased Akt activity and an EMT phenotype, including a switch from TWIST2 to TWIST1 expression [115]. The same study found that the expression of LMNA reversed EMT via the inhibition of MAPK signals and preserved drug sensitivity.…”