GRP78 blockade overcomes acquired resistance to EGFR-tyrosine kinase inhibitors in non-small cell lung cancer

Life Sci. 2024 Jul 1:348:122681. doi: 10.1016/j.lfs.2024.122681. Epub 2024 Apr 30.

Abstract

Aims: While significant upregulation of GRP78 has been documented in lung cancer patients, its association with resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) remains underexamined. Our study aimed to elucidate the functional importance of GRP78 in acquired resistance to EGFR-TKIs in non-small cell lung cancer (NSCLC) and to evaluate its potential as a therapeutic target.

Main methods: Immunoblot analysis or flow cytometry was employed to assess several markers for endoplasmic reticulum (ER) stress and apoptosis. Ru(II) complex I and HA15, two known GRP78 inhibitors, were used to evaluate the functional role of GRP78. A Xenograft assay was performed to evaluate the in vivo anti-cancer effects of the GRP78 inhibitors.

Key findings: We validated a significant increase in GRP78 protein levels in HCC827-GR, H1993-GR, and H1993-ER cells. The EGFR-TKI-resistant cells overexpressing GRP78 exhibited significantly higher cell proliferation rates than did their parental counterparts. Notably, GRP78 inhibition resulted in a more profound anti-proliferative and apoptotic response via heightened ER stress and subsequent reactive oxygen species (ROS) production in EGFR-TKI-resistant cell lines compared with their parental cells. In xenograft models implanted with HCC827-GR, both Ru(II) complex I and HA15 significantly suppressed tumor growth and reduced tumor weight. Additionally, we confirmed that GRP78 plays a critical role in the proliferation of H1975, an EGFR-TKI-resistant T790M-mutant cell line, relative to other NSCLC cell lines.

Significance: Our findings strongly support targeting of GRP78 as a promising therapeutic strategy for NSCLC patients with acquired resistance to EGFR-TKIs.

Keywords: EGFR-TKI resistance; ER stress; GRP78; NSCLC; Therapeutic target.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Carcinoma, Non-Small-Cell Lung* / drug therapy
  • Carcinoma, Non-Small-Cell Lung* / metabolism
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Drug Resistance, Neoplasm* / drug effects
  • Endoplasmic Reticulum Chaperone BiP*
  • Endoplasmic Reticulum Stress / drug effects
  • ErbB Receptors* / antagonists & inhibitors
  • ErbB Receptors* / metabolism
  • Female
  • Heat-Shock Proteins* / metabolism
  • Humans
  • Lung Neoplasms* / drug therapy
  • Lung Neoplasms* / metabolism
  • Lung Neoplasms* / pathology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Reactive Oxygen Species / metabolism
  • Tyrosine Kinase Inhibitors* / pharmacology
  • Xenograft Model Antitumor Assays*

Substances

  • EGFR protein, human
  • Endoplasmic Reticulum Chaperone BiP
  • ErbB Receptors
  • Heat-Shock Proteins
  • HSPA5 protein, human
  • Hspa5 protein, mouse
  • Reactive Oxygen Species
  • Tyrosine Kinase Inhibitors