Limits...
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.

Show MeSH

Related in: MedlinePlus

The therapy-induced secretome of sensitive cells overlaps significantly in melanoma and lung adenocarcinoma cells and appears after gene expression changes enriched for transcriptional regulatorsa–b, GO analysis (revigo.irb.hr) of gene expression changes after 6h of vemurafenib treatment of A375 cells with (a) spatial representation of enriched GO terms and (b) the molecular functions significantly affected. c, Heat map, representing the expression levels of commonly up- and down-regulated genes in vemurafenib-treated A375-derived xenograft tumours (5 days) and A375 cells in vitro (48h). d, Principal component analysis of vemurafenib-sensitive Colo800 and UACC62 melanoma cells and crizotinib-sensitive H3122 lung adenocarcinoma cells treated in vitro with vehicle or vemurafenib or crizotinib for 48h. e, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. g, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) down-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. P values shown were calculated using a Hypergeometric probability test.
© Copyright Policy - permissions-link
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4507807&req=5

Figure 7: The therapy-induced secretome of sensitive cells overlaps significantly in melanoma and lung adenocarcinoma cells and appears after gene expression changes enriched for transcriptional regulatorsa–b, GO analysis (revigo.irb.hr) of gene expression changes after 6h of vemurafenib treatment of A375 cells with (a) spatial representation of enriched GO terms and (b) the molecular functions significantly affected. c, Heat map, representing the expression levels of commonly up- and down-regulated genes in vemurafenib-treated A375-derived xenograft tumours (5 days) and A375 cells in vitro (48h). d, Principal component analysis of vemurafenib-sensitive Colo800 and UACC62 melanoma cells and crizotinib-sensitive H3122 lung adenocarcinoma cells treated in vitro with vehicle or vemurafenib or crizotinib for 48h. e, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. g, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) down-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. P values shown were calculated using a Hypergeometric probability test.

Mentions: To identify relevant components and regulators of the reactive secretome, we analysed gene expression changes in sensitive A375 melanoma cells at different time points after vemurafenib exposure in vitro. After 6h on vemurafenib, 473 genes showed altered expression and pathway analysis revealed that these genes were enriched for transcriptional regulators (Fig. 3a, b, Extended Data Fig. 3a, b, Supplementary Table 1). After 48h, more than one third of the transcriptome was differentially expressed (>5000 genes; 405 genes encoding for proteins in the extracellular region GO:0005576), significantly overlapping with the gene expression changes of A375 tumours in vivo after 5 days of vemurafenib treatment (Fig. 3a, b, Extended Data Fig. 3c). Similar extensive gene expression changes were observed in Colo800 and UACC62 melanoma cells treated with vemurafenib and H3122 lung adenocarcinoma cells treated with crizotinib (Extended Data Fig 3d). Despite different cell lineages, different oncogenic drivers, and different targeted therapies we observed a significant overlap between the secretome of melanoma and lung adenocarcinoma cells (p<9.11E–5) (Extended Data Fig. 3e–h, Supplementary Table 1). Furthermore, changes in the secretome of vemurafenib-sensitive melanoma cells coincided with changes in the immune cell composition (Extended Data Fig. 4a, b), and with changes of soluble mediators derived from murine stromal cells such as IGF1 and HGF (Extended Data Fig. 4c, d). These data indicate a therapy-induced secretome (TIS), a response that consists of many up- and down-regulated secreted factors, permeates the regressing tumour microenvironment and stimulates cancer cells, likely also stromal cells.


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)

The therapy-induced secretome of sensitive cells overlaps significantly in melanoma and lung adenocarcinoma cells and appears after gene expression changes enriched for transcriptional regulatorsa–b, GO analysis (revigo.irb.hr) of gene expression changes after 6h of vemurafenib treatment of A375 cells with (a) spatial representation of enriched GO terms and (b) the molecular functions significantly affected. c, Heat map, representing the expression levels of commonly up- and down-regulated genes in vemurafenib-treated A375-derived xenograft tumours (5 days) and A375 cells in vitro (48h). d, Principal component analysis of vemurafenib-sensitive Colo800 and UACC62 melanoma cells and crizotinib-sensitive H3122 lung adenocarcinoma cells treated in vitro with vehicle or vemurafenib or crizotinib for 48h. e, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. g, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) down-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. P values shown were calculated using a Hypergeometric probability test.
© Copyright Policy - permissions-link
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4507807&req=5

Figure 7: The therapy-induced secretome of sensitive cells overlaps significantly in melanoma and lung adenocarcinoma cells and appears after gene expression changes enriched for transcriptional regulatorsa–b, GO analysis (revigo.irb.hr) of gene expression changes after 6h of vemurafenib treatment of A375 cells with (a) spatial representation of enriched GO terms and (b) the molecular functions significantly affected. c, Heat map, representing the expression levels of commonly up- and down-regulated genes in vemurafenib-treated A375-derived xenograft tumours (5 days) and A375 cells in vitro (48h). d, Principal component analysis of vemurafenib-sensitive Colo800 and UACC62 melanoma cells and crizotinib-sensitive H3122 lung adenocarcinoma cells treated in vitro with vehicle or vemurafenib or crizotinib for 48h. e, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. g, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) down-regulated after 48h of vemurafenib treatment in A375, Colo800, and UACC62 melanoma cell lines. f, Venn Diagram indicating the overlap of genes in the extracellular region (GO:0005576) up-regulated after 48h of vemurafenib treatment in at least 2/3 melanoma models and after 48h of crizotinib treatment in the H3122 lung adenocarcinoma cell line. P values shown were calculated using a Hypergeometric probability test.
Mentions: To identify relevant components and regulators of the reactive secretome, we analysed gene expression changes in sensitive A375 melanoma cells at different time points after vemurafenib exposure in vitro. After 6h on vemurafenib, 473 genes showed altered expression and pathway analysis revealed that these genes were enriched for transcriptional regulators (Fig. 3a, b, Extended Data Fig. 3a, b, Supplementary Table 1). After 48h, more than one third of the transcriptome was differentially expressed (>5000 genes; 405 genes encoding for proteins in the extracellular region GO:0005576), significantly overlapping with the gene expression changes of A375 tumours in vivo after 5 days of vemurafenib treatment (Fig. 3a, b, Extended Data Fig. 3c). Similar extensive gene expression changes were observed in Colo800 and UACC62 melanoma cells treated with vemurafenib and H3122 lung adenocarcinoma cells treated with crizotinib (Extended Data Fig 3d). Despite different cell lineages, different oncogenic drivers, and different targeted therapies we observed a significant overlap between the secretome of melanoma and lung adenocarcinoma cells (p<9.11E–5) (Extended Data Fig. 3e–h, Supplementary Table 1). Furthermore, changes in the secretome of vemurafenib-sensitive melanoma cells coincided with changes in the immune cell composition (Extended Data Fig. 4a, b), and with changes of soluble mediators derived from murine stromal cells such as IGF1 and HGF (Extended Data Fig. 4c, d). These data indicate a therapy-induced secretome (TIS), a response that consists of many up- and down-regulated secreted factors, permeates the regressing tumour microenvironment and stimulates cancer cells, likely also stromal cells.

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