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Published on: April 6, 2016
TP63 drives TKI resistance in EGFR-mutant lung cancer via ferroptosis inhibition
Kehan Li1, Jiane Liu2, Wenjuan Wang3
1Department of Genetics and Cell Biology, School of Basic Medicine, Qingdao University, Qingdao, China.
Background:
The emergence of acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) presents a significant obstacle in the therapeutic strategy for EGFR-mutant non-small cell lung cancer (NSCLC). While secondary mutations explain some cases, adaptive resistance mechanisms involving lineage plasticity and metabolic rewiring are increasingly recognized but poorly understood. This study aimed to investigate the role of the transcription factor TP63 in mediating adaptive resistance to EGFR-TKIs, with a focus on its potential regulation of ferroptosis.
Methods:
To investigate these mechanisms, we generated osimertinib-resistant derivatives of EGFR-mutant PC9 and HCC827 cell lines. We utilized short hairpin RNA (shRNA) interference to silence TP63 and performed functional assays including cell viability, colony formation, wound-healing, and transwell invasion assays. Mechanistic insights were gained through integrated RNA sequencing (RNA-seq), immunofluorescence, and lipid peroxidation assays using BODIPY 581/591 C11.
Results:
We identified the transcription factor TP63 as a master regulator of adaptive resistance, showing marked upregulation in resistant cells. Although TP63 depletion did not affect basal cell viability, its loss significantly restored sensitivity to osimertinib, suppressing proliferation, clonogenic growth, and invasive potential. Transcriptomic and biochemical analyses revealed that TP63 maintains redox homeostasis by transcriptionally activating GPX4 and antioxidant gene networks. Consequently, TP63 silencing collapsed this defense system, leading to the accumulation of lipid peroxides and inducing ferroptotic cell death under drug pressure.
Conclusions:
Clinically, high TP63 expression correlates with reduced overall survival in EGFR-mutant NSCLC patients. These findings define a critical TP63-GPX4 ferroptosis axis that safeguards tumor cell survival during therapeutic stress, highlighting ferroptosis induction as a potential therapeutic strategy to overcome EGFR-TKI resistance.
Insights
The transcription factor TP63 drives adaptive resistance to EGFR tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) by activating the ferroptosis suppressor GPX4. Inhibiting TP63 re-sensitizes tumors to TKIs.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) is a major challenge in treating EGFR-mutant non-small cell lung cancer (NSCLC).
- Adaptive resistance mechanisms, including lineage plasticity and metabolic alterations, are implicated but not fully understood.
- The role of transcription factor TP63 in mediating EGFR-TKI resistance and its connection to ferroptosis remain largely unexplored.
Purpose of the Study:
- To investigate the role of the transcription factor TP63 in adaptive resistance to EGFR-TKIs in NSCLC.
- To elucidate the mechanisms by which TP63 influences tumor cell survival under TKI treatment.
- To explore the potential of targeting the TP63-ferroptosis axis for overcoming therapeutic resistance.
Main Methods:
- Generation of osimertinib-resistant EGFR-mutant NSCLC cell lines (PC9 and HCC827).
- Utilized short hairpin RNA (shRNA) to silence TP63, followed by functional assays (cell viability, colony formation, migration, invasion).
- Integrated transcriptomic (RNA-seq), immunofluorescence, and lipid peroxidation assays to analyze molecular mechanisms.
Main Results:
- TP63 was identified as a key regulator of adaptive resistance, with significantly upregulated expression in resistant cells.
- TP63 depletion restored sensitivity to osimertinib, inhibiting proliferation and invasion, despite not affecting basal cell viability.
- TP63 maintains redox homeostasis by upregulating GPX4 and antioxidant genes, and its silencing induces ferroptosis by causing lipid peroxide accumulation.
Conclusions:
- High TP63 expression correlates with poorer survival in EGFR-mutant NSCLC patients.
- A critical TP63-GPX4 ferroptosis axis was defined, which protects tumor cells during EGFR-TKI therapy.
- Targeting ferroptosis induction presents a promising strategy to overcome acquired resistance to EGFR-TKIs in NSCLC.
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