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Therapy-induced tumour secretomes promote resistance and tumour progression.

Obenauf AC, Zou Y, Ji AL, Vanharanta S, Shu W, Shi H, Kong X, Bosenberg MC, Wiesner T, Rosen N, Lo RS, Massagué J - Nature (2015)

Bottom Line: Drug resistance invariably limits the clinical efficacy of targeted therapy with kinase inhibitors against cancer.Here we show that targeted therapy with BRAF, ALK or EGFR kinase inhibitors induces a complex network of secreted signals in drug-stressed human and mouse melanoma and human lung adenocarcinoma cells.The tumour-promoting secretome of melanoma cells treated with the kinase inhibitor vemurafenib is driven by downregulation of the transcription factor FRA1.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA.

ABSTRACT
Drug resistance invariably limits the clinical efficacy of targeted therapy with kinase inhibitors against cancer. Here we show that targeted therapy with BRAF, ALK or EGFR kinase inhibitors induces a complex network of secreted signals in drug-stressed human and mouse melanoma and human lung adenocarcinoma cells. This therapy-induced secretome stimulates the outgrowth, dissemination and metastasis of drug-resistant cancer cell clones and supports the survival of drug-sensitive cancer cells, contributing to incomplete tumour regression. The tumour-promoting secretome of melanoma cells treated with the kinase inhibitor vemurafenib is driven by downregulation of the transcription factor FRA1. In situ transcriptome analysis of drug-resistant melanoma cells responding to the regressing tumour microenvironment revealed hyperactivation of several signalling pathways, most prominently the AKT pathway. Dual inhibition of RAF and the PI(3)K/AKT/mTOR intracellular signalling pathways blunted the outgrowth of the drug-resistant cell population in BRAF mutant human melanoma, suggesting this combination therapy as a strategy against tumour relapse. Thus, therapeutic inhibition of oncogenic drivers induces vast secretome changes in drug-sensitive cancer cells, paradoxically establishing a tumour microenvironment that supports the expansion of drug-resistant clones, but is susceptible to combination therapy.

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Targeted therapy induces down-regulation of FRA1 in drug-sensitive tumour cellsa, List of transcription factors predicted to regulate the vemurafenib-induced reactive secretome in A375 cells, and a heat map of the corresponding transcription factor gene expression levels in these cells. Red represents high, yellow medium and blue low relative expression on the colour scale. b, Immunoblotting of phosphorylated and total FRA1 protein levels in A375 and YUMM1.7 melanoma cell lines treated with vemurafenib for 24h. c, Relative mRNA levels of FRA1 in H3122 cells treated with crizotinib (500nM) and HCC827 treated with erlotinib (10nM) at different time points (n = 4 technical replicates) d, Relative mRNA levels of FRA1 in A375R cells treated with vemurafenib at different time points (n = 4 technical replicates). e, Immunofluorescence staining of FRA1 (red) and DAPI (blue) in biopsies from melanoma patients before and after vemurafenib treatment (clinical information can be found in Extended Data Table 1).
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Figure 9: Targeted therapy induces down-regulation of FRA1 in drug-sensitive tumour cellsa, List of transcription factors predicted to regulate the vemurafenib-induced reactive secretome in A375 cells, and a heat map of the corresponding transcription factor gene expression levels in these cells. Red represents high, yellow medium and blue low relative expression on the colour scale. b, Immunoblotting of phosphorylated and total FRA1 protein levels in A375 and YUMM1.7 melanoma cell lines treated with vemurafenib for 24h. c, Relative mRNA levels of FRA1 in H3122 cells treated with crizotinib (500nM) and HCC827 treated with erlotinib (10nM) at different time points (n = 4 technical replicates) d, Relative mRNA levels of FRA1 in A375R cells treated with vemurafenib at different time points (n = 4 technical replicates). e, Immunofluorescence staining of FRA1 (red) and DAPI (blue) in biopsies from melanoma patients before and after vemurafenib treatment (clinical information can be found in Extended Data Table 1).

Mentions: To identify molecular drivers of the A375-TIS in response to vemurafenib, we integrated the data of differentially expressed transcription factors after 6h of vemurafenib treatment with the transcription factor binding motifs that were enriched at the promoters of differentially expressed genes in the secretome after 48h (Fig. 3a, b). This analysis highlighted FRA1 (FOSL1), a member of the AP1 transcription factor complex and effector of the ERK pathway27, as one of the putative upstream regulators of the TIS (Extended Data Fig. 5a). FRA1 was down-regulated in all drug-sensitive cells, but not in resistant cells, treated with vemurafenib, crizotinib, and erlotinib (Fig. 3c, d, Extended Data Fig. 5b–d). Biopsies from melanoma patients early during RAFi treatment confirmed vemurafenib-induced FRA1 down-regulation in clinical samples (Fig. 3e, Extended Data Fig. 5e, Extended Data Table 1).


Therapy-induced tumour secretomes promote resistance and tumour progression.

Obenauf AC, Zou Y, Ji AL, Vanharanta S, Shu W, Shi H, Kong X, Bosenberg MC, Wiesner T, Rosen N, Lo RS, Massagué J - Nature (2015)

Targeted therapy induces down-regulation of FRA1 in drug-sensitive tumour cellsa, List of transcription factors predicted to regulate the vemurafenib-induced reactive secretome in A375 cells, and a heat map of the corresponding transcription factor gene expression levels in these cells. Red represents high, yellow medium and blue low relative expression on the colour scale. b, Immunoblotting of phosphorylated and total FRA1 protein levels in A375 and YUMM1.7 melanoma cell lines treated with vemurafenib for 24h. c, Relative mRNA levels of FRA1 in H3122 cells treated with crizotinib (500nM) and HCC827 treated with erlotinib (10nM) at different time points (n = 4 technical replicates) d, Relative mRNA levels of FRA1 in A375R cells treated with vemurafenib at different time points (n = 4 technical replicates). e, Immunofluorescence staining of FRA1 (red) and DAPI (blue) in biopsies from melanoma patients before and after vemurafenib treatment (clinical information can be found in Extended Data Table 1).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4507807&req=5

Figure 9: Targeted therapy induces down-regulation of FRA1 in drug-sensitive tumour cellsa, List of transcription factors predicted to regulate the vemurafenib-induced reactive secretome in A375 cells, and a heat map of the corresponding transcription factor gene expression levels in these cells. Red represents high, yellow medium and blue low relative expression on the colour scale. b, Immunoblotting of phosphorylated and total FRA1 protein levels in A375 and YUMM1.7 melanoma cell lines treated with vemurafenib for 24h. c, Relative mRNA levels of FRA1 in H3122 cells treated with crizotinib (500nM) and HCC827 treated with erlotinib (10nM) at different time points (n = 4 technical replicates) d, Relative mRNA levels of FRA1 in A375R cells treated with vemurafenib at different time points (n = 4 technical replicates). e, Immunofluorescence staining of FRA1 (red) and DAPI (blue) in biopsies from melanoma patients before and after vemurafenib treatment (clinical information can be found in Extended Data Table 1).
Mentions: To identify molecular drivers of the A375-TIS in response to vemurafenib, we integrated the data of differentially expressed transcription factors after 6h of vemurafenib treatment with the transcription factor binding motifs that were enriched at the promoters of differentially expressed genes in the secretome after 48h (Fig. 3a, b). This analysis highlighted FRA1 (FOSL1), a member of the AP1 transcription factor complex and effector of the ERK pathway27, as one of the putative upstream regulators of the TIS (Extended Data Fig. 5a). FRA1 was down-regulated in all drug-sensitive cells, but not in resistant cells, treated with vemurafenib, crizotinib, and erlotinib (Fig. 3c, d, Extended Data Fig. 5b–d). Biopsies from melanoma patients early during RAFi treatment confirmed vemurafenib-induced FRA1 down-regulation in clinical samples (Fig. 3e, Extended Data Fig. 5e, Extended Data Table 1).

Bottom Line: Drug resistance invariably limits the clinical efficacy of targeted therapy with kinase inhibitors against cancer.Here we show that targeted therapy with BRAF, ALK or EGFR kinase inhibitors induces a complex network of secreted signals in drug-stressed human and mouse melanoma and human lung adenocarcinoma cells.The tumour-promoting secretome of melanoma cells treated with the kinase inhibitor vemurafenib is driven by downregulation of the transcription factor FRA1.

View Article: PubMed Central - PubMed

Affiliation: Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA.

ABSTRACT
Drug resistance invariably limits the clinical efficacy of targeted therapy with kinase inhibitors against cancer. Here we show that targeted therapy with BRAF, ALK or EGFR kinase inhibitors induces a complex network of secreted signals in drug-stressed human and mouse melanoma and human lung adenocarcinoma cells. This therapy-induced secretome stimulates the outgrowth, dissemination and metastasis of drug-resistant cancer cell clones and supports the survival of drug-sensitive cancer cells, contributing to incomplete tumour regression. The tumour-promoting secretome of melanoma cells treated with the kinase inhibitor vemurafenib is driven by downregulation of the transcription factor FRA1. In situ transcriptome analysis of drug-resistant melanoma cells responding to the regressing tumour microenvironment revealed hyperactivation of several signalling pathways, most prominently the AKT pathway. Dual inhibition of RAF and the PI(3)K/AKT/mTOR intracellular signalling pathways blunted the outgrowth of the drug-resistant cell population in BRAF mutant human melanoma, suggesting this combination therapy as a strategy against tumour relapse. Thus, therapeutic inhibition of oncogenic drivers induces vast secretome changes in drug-sensitive cancer cells, paradoxically establishing a tumour microenvironment that supports the expansion of drug-resistant clones, but is susceptible to combination therapy.

Show MeSH
Related in: MedlinePlus