RÉSUMÉ
Tumors with mutant RAS are often dependent on extracellular signal-regulated kinase (ERK) signaling for growth; however, MEK inhibitors have only marginal antitumor activity in these tumors. MEK inhibitors relieve ERK-dependent feedback inhibition of RAF and cause induction of MEK phosphorylation. We have now identified a MEK inhibitor, CH5126766 (RO5126766), that has the unique property of inhibiting RAF kinase as well. CH5126766 binding causes MEK to adopt a conformation in which it cannot be phosphorylated by and released from RAF. This results in formation of a stable MEK/RAF complex and inhibition of RAF kinase. Consistent with this mechanism, this drug does not induce MEK phosphorylation. CH5126766 inhibits ERK signaling output more effectively than a standard MEK inhibitor that induces MEK phosphorylation and has potent antitumor activity as well. These results suggest that relief of RAF feedback limits pathway inhibition by standard MEK inhibitors. CH5126766 represents a new type of MEK inhibitor that causes MEK to become a dominant-negative inhibitor of RAF and that, in doing so, may have enhanced therapeutic activity in ERK-dependent tumors with mutant RAS.
Sujet(s)
Antinéoplasiques/pharmacologie , Coumarines/pharmacologie , MAP Kinase Kinase 1/antagonistes et inhibiteurs , Système de signalisation des MAP kinases/effets des médicaments et des substances chimiques , Protéines proto-oncogènes B-raf/métabolisme , Régulation allostérique , Animaux , Lignée cellulaire tumorale , Activation enzymatique/effets des médicaments et des substances chimiques , Extracellular Signal-Regulated MAP Kinases/métabolisme , Rétrocontrôle physiologique/effets des médicaments et des substances chimiques , Femelle , Humains , MAP Kinase Kinase 1/composition chimique , MAP Kinase Kinase 1/métabolisme , Souris , Souris de lignée BALB C , Souris nude , Phosphorylation , Liaison aux protéines , Maturation post-traductionnelle des protéines , Protéines proto-oncogènes/génétique , Protéines proto-oncogènes B-raf/composition chimique , Protéines proto-oncogènes c-raf/composition chimique , Protéines proto-oncogènes c-raf/métabolisme , Protéines proto-oncogènes p21(ras) , Tests d'activité antitumorale sur modèle de xénogreffe , Protéines G ras/génétiqueRÉSUMÉ
BRAF(V600E) drives tumors by dysregulating ERK signaling. In these tumors, we show that high levels of ERK-dependent negative feedback potently suppress ligand-dependent mitogenic signaling and Ras function. BRAF(V600E) activation is Ras independent and it signals as a RAF-inhibitor-sensitive monomer. RAF inhibitors potently inhibit RAF monomers and ERK signaling, causing relief of ERK-dependent feedback, reactivation of ligand-dependent signal transduction, increased Ras-GTP, and generation of RAF-inhibitor-resistant RAF dimers. This results in a rebound in ERK activity and culminates in a new steady state, wherein ERK signaling is elevated compared to its initial nadir after RAF inhibition. In this state, ERK signaling is RAF inhibitor resistant, and MEK inhibitor sensitive, and combined inhibition results in enhancement of ERK pathway inhibition and antitumor activity.
Sujet(s)
Système de signalisation des MAP kinases/effets des médicaments et des substances chimiques , Mélanome/génétique , Protéines proto-oncogènes B-raf/génétique , Protéines G ras/antagonistes et inhibiteurs , Lignée cellulaire tumorale , Facteur de croissance épidermique/métabolisme , Extracellular Signal-Regulated MAP Kinases/génétique , Extracellular Signal-Regulated MAP Kinases/métabolisme , Extracellular Signal-Regulated MAP Kinases/physiologie , Facteurs de croissance fibroblastique/métabolisme , Régulation de l'expression des gènes tumoraux , Facteur de croissance des hépatocytes/métabolisme , Humains , Indoles/pharmacologie , Protéines et peptides de signalisation intracellulaire/génétique , Protéines et peptides de signalisation intracellulaire/métabolisme , Protéines et peptides de signalisation intracellulaire/physiologie , Ligands , Mélanome/métabolisme , Protéines membranaires , Neurégulines/métabolisme , Protéines proto-oncogènes B-raf/métabolisme , Protéines proto-oncogènes B-raf/physiologie , Récepteur facteur croissance/métabolisme , Sulfonamides/pharmacologie , Vémurafénib , Protéines G ras/métabolisme , Protéines G ras/physiologieRÉSUMÉ
Tumors with mutant BRAF and some with mutant RAS are dependent upon ERK signaling for proliferation, and their growth is suppressed by MAPK/ERK kinase (MEK) inhibitors. In contrast, tumor cells with human EGF receptor (HER) kinase activation proliferate in a MEK-independent manner. These findings have led to the development of RAF and MEK inhibitors as anticancer agents. Like MEK inhibitors, the RAF inhibitor PLX4032 inhibits the proliferation of BRAF(V600E) tumor cells but not that of HER kinase-dependent tumors. However, tumors with RAS mutation that are sensitive to MEK inhibition are insensitive to PLX4032. MEK inhibitors inhibit ERK phosphorylation in all normal and tumor cells, whereas PLX4032 inhibits ERK signaling only in tumor cells expressing BRAF(V600E). In contrast, the drug activates MEK and ERK phosphorylation in cells with wild-type BRAF. In BRAF(V600E) tumor cells, MEK and RAF inhibitors affect the expression of a common set of genes. PLX4032 inhibits ERK signaling output in mutant BRAF cells, whereas it transiently activates the expression of these genes in tumor cells with wild-type RAF. Thus, PLX4032 inhibits ERK signaling output in a mutant BRAF-selective manner. These data explain why the drug selectively inhibits the growth of mutant BRAF tumors and suggest that it will not cause toxicity resulting from the inhibition of ERK signaling in normal cells. This selectivity may lead to a broader therapeutic index and help explain the greater antitumor activity observed with this drug than with MEK inhibitors.