COT drives resistance to RAF inhibition through MAP kinase pathway reactivation

• Published in: Nature (London) Vol. 468; no. 7326; pp. 968 - 972
• Main Authors: Johannessen, Cory M., Boehm, Jesse S., Kim, So Young, Thomas, Sapana R., Wardwell, Leslie, Johnson, Laura A., Emery, Caroline M., Stransky, Nicolas, Cogdill, Alexandria P., Barretina, Jordi, Caponigro, Giordano, Hieronymus, Haley, Murray, Ryan R., Salehi-Ashtiani, Kourosh, Hill, David E., Vidal, Marc, Zhao, Jean J., Yang, Xiaoping, Alkan, Ozan, Kim, Sungjoon, Harris, Jennifer L., Wilson, Christopher J., Myer, Vic E., Finan, Peter M., Root, David E., Roberts, Thomas M., Golub, Todd, Flaherty, Keith T., Dummer, Reinhard, Weber, Barbara L., Sellers, William R., Schlegel, Robert, Wargo, Jennifer A., Hahn, William C., Garraway, Levi A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.12.2010; Nature Publishing Group
• Subjects: 631/67/1059/2326; 692/699/67/1813/1634; Allosteric Regulation; Antineoplastic agents; Biochemistry; Biological and medical sciences; Cancer; Cancer cells; Cell Line, Tumor; Cell lines; Clinical Trials as Topic; Drug Resistance, Neoplasm - drug effects; Drug Resistance, Neoplasm - genetics; Enzyme Activation - drug effects; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gene Library; General aspects; Genes; Humanities and Social Sciences; Humans; Indoles - pharmacology; Indoles - therapeutic use; Kinases; letter; MAP Kinase Kinase Kinases - genetics; MAP Kinase Kinase Kinases - metabolism; MAP Kinase Signaling System; Medical sciences; Melanoma; Melanoma - drug therapy; Melanoma - enzymology; Melanoma - genetics; Melanoma - metabolism; Mitogen-Activated Protein Kinase Kinases - antagonists & inhibitors; Mitogen-Activated Protein Kinase Kinases - metabolism; Mitogen-Activated Protein Kinases - metabolism; multidisciplinary; Mutation; Open Reading Frames - genetics; Pharmacology. Drug treatments; Properties; Protein Kinase Inhibitors - pharmacology; Protein Kinase Inhibitors - therapeutic use; Protein kinases; Protein research; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins B-raf - antagonists & inhibitors; Proto-Oncogene Proteins B-raf - chemistry; Proto-Oncogene Proteins B-raf - genetics; Proto-Oncogene Proteins B-raf - metabolism; Proto-Oncogene Proteins c-raf - genetics; Proto-Oncogene Proteins c-raf - metabolism; Science; Science (multidisciplinary); Sulfonamides - pharmacology; Sulfonamides - therapeutic use; Vemurafenib; United States; Antineoplastic agent; Human; Treatment resistance; Enzyme; Transferases; Enzyme inhibitor; Mitogen-activated protein kinase; Malignant tumor; Open reading frame; Cancerology; Cell line; Malignant melanoma; Genetics; Cancer
• ISSN: 0028-0836; 1476-4687; 1476-4687
• DOI: 10.1038/nature09627

Drug-resistance mechanism in melanoma Clinical trials in melanoma patients carrying B-RAF gene mutations have shown promising results with the B-RAF kinase inhibitor PLX4032, but many patients go on to become resistant. Two papers now uncover possible mechanisms for this resistance. Nazarian et al . report that melanomas can acquire resistance due to mutations of N-RAS or increased expression of PDGFRβ , and Johannessen et al . report resistance due to upregulation of MAP3K8/COT. Each of these mechanisms seems to apply to some patients in the recent PLX4032 trial, yet surprisingly, no secondary B-RAF mutations were observed. Recent data from early clinical trials in melanoma patients carrying mutations in the B-RAF gene have shown promising results with the B-RAF kinase inhibitor PLX4032; however, many patients eventually develop resistance to this treatment. Two papers now uncover possible mechanisms of resistance to PLX4032. One paper shows that upregulation of MAP3K8 (which encodes COT) can confer resistance of melanoma cells to B-RAF inhibitors, whereas another paper found that melanomas can acquire resistance due to mutations of N-RAS or increased expression of PDGFRβ. Each of these resistance mechanisms seems to apply to at least some patients on recent PLX4032 trial, whereas, surprisingly, so far no secondary B-RAF mutations have been observed. Oncogenic mutations in the serine/threonine kinase B-RAF (also known as BRAF) are found in 50–70% of malignant melanomas 1 . Pre-clinical studies have demonstrated that the B-RAF(V600E) mutation predicts a dependency on the mitogen-activated protein kinase (MAPK) signalling cascade in melanoma 2 , 3 , 4 , 5 , 6 —an observation that has been validated by the success of RAF and MEK inhibitors in clinical trials 7 , 8 , 9 . However, clinical responses to targeted anticancer therapeutics are frequently confounded by de novo or acquired resistance 10 , 11 , 12 . Identification of resistance mechanisms in a manner that elucidates alternative ‘druggable’ targets may inform effective long-term treatment strategies 13 . Here we expressed ∼600 kinase and kinase-related open reading frames (ORFs) in parallel to interrogate resistance to a selective RAF kinase inhibitor. We identified MAP3K8 (the gene encoding COT/Tpl2) as a MAPK pathway agonist that drives resistance to RAF inhibition in B-RAF(V600E) cell lines. COT activates ERK primarily through MEK-dependent mechanisms that do not require RAF signalling. Moreover, COT expression is associated with de novo resistance in B-RAF(V600E) cultured cell lines and acquired resistance in melanoma cells and tissue obtained from relapsing patients following treatment with MEK or RAF inhibitors. We further identify combinatorial MAPK pathway inhibition or targeting of COT kinase activity as possible therapeutic strategies for reducing MAPK pathway activation in this setting. Together, these results provide new insights into resistance mechanisms involving the MAPK pathway and articulate an integrative approach through which high-throughput functional screens may inform the development of novel therapeutic strategies.