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AXL-Mediated Drug Resistance in ALK-Rearranged NSCLC Enhanced by GAS6 From Macrophages and MMP11 Positive Fibroblasts
Takahiro Utsumi1,2, Hayato Mizuta1,3, Yosuke Seto1
1Division of Experimental Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo, Japan.
Abstract:
Anaplastic lymphoma kinase (ALK) rearranged non-small cell lung cancer (NSCLC) shows marked tumor shrinkage by ALK-tyrosine kinase inhibitors (TKIs). However, tumors almost inevitably relapse owing to the development of acquired resistance. Resistance mechanisms include secondary ALK mutations and the activation of bypass pathways, such as cMET, cKIT, or EGFR, though some remain unknown. In this study, we analyzed alectinib-resistant patient samples and identified a significant increase in AXL expression in the tumor, and a high level of GAS6, the ligand for AXL, in the pleural effusion. AXL-overexpressing H3122 ALK-rearranged NSCLC cells exhibited partial resistance to alectinib, which was enhanced by GAS6 supplementation but could be overcome by the ALK/AXL inhibitor gilteritinib. Moreover, GAS6-overexpressing NIH3T3 cells and AXL-expressing H3122 cells were subcutaneously injected into the left and right sides of nude mice simultaneously, followed by alectinib treatment. The supply of GAS6 from NIH3T3 may have accelerated tumor relapse under alectinib treatment. However, even without GAS6-overexpressing NIH3T3, AXL-overexpressing H3122 tumor relapsed within 1 month possibly due to increased host mouse Gas6 expression. Single-cell RNA sequencing revealed that specific cancer-associated fibroblasts (CAFs) and a subset of tumor-associated macrophages (TAMs) are the primary sources of Gas6 in the tumor microenvironment (TME). During alectinib treatment, TAMs increased their infiltration into the TME, whereas CAFs altered their expression patterns, substantially upregulating Mmp11. These findings suggest that AXL expression in resistant cancer cells, combined with increased Gas6 production in the TME, contributes to enhanced ALK-TKI resistance.
Insights
Resistance to ALK-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) is linked to increased AXL expression and GAS6 ligand. Targeting both ALK and AXL may overcome this resistance.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Anaplastic lymphoma kinase (ALK) rearranged non-small cell lung cancer (NSCLC) initially responds well to ALK-tyrosine kinase inhibitors (TKIs).
- Acquired resistance to ALK-TKIs is a major clinical challenge, often involving secondary mutations or bypass signaling pathways.
- The precise mechanisms driving resistance, particularly those involving unknown factors, require further elucidation.
Purpose of the Study:
- To investigate the mechanisms of acquired resistance to alectinib in ALK-rearranged NSCLC.
- To identify novel molecular players involved in treatment failure and tumor relapse.
- To explore potential therapeutic strategies to overcome alectinib resistance.
Main Methods:
- Analysis of alectinib-resistant patient samples.
- In vitro studies using AXL-overexpressing NSCLC cell lines with and without GAS6 supplementation.
- In vivo mouse models with co-injected cancer cells and xenografts.
- Single-cell RNA sequencing of tumor microenvironment components.
Main Results:
- Increased AXL expression in resistant tumors and elevated GAS6 levels in pleural effusion were observed.
- AXL-overexpressing cells showed partial resistance to alectinib, exacerbated by GAS6 but sensitive to gilteritinib (ALK/AXL inhibitor).
- In vivo models demonstrated accelerated relapse with GAS6 supply and identified cancer-associated fibroblasts and tumor-associated macrophages as key Gas6 sources in the tumor microenvironment.
Conclusions:
- AXL expression in cancer cells and increased GAS6 production within the tumor microenvironment contribute significantly to acquired resistance against ALK-TKIs.
- Targeting the AXL pathway, in combination with ALK inhibition, represents a promising strategy to overcome resistance in ALK-rearranged NSCLC.
- Understanding the role of tumor microenvironment components like CAFs and TAMs in resistance is crucial for developing effective therapies.
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