title: The CDK4/6 inhibitor LY2835219 overcomes vemurafenib resistance resulting from MAPK reactivation and cyclin D1 upregulation.
abstract: B-RAF selective inhibitors, including vemurafenib, were recently developed as effective therapies for melanoma patients with B-RAF V600E mutation. However, most patients treated with vemurafenib eventually develop resistance largely due to reactivation of MAPK signaling. Inhibitors of MAPK signaling, including MEK1/2 inhibitor trametinib, failed to show significant clinical benefit in patients with acquired resistance to vemurafenib. Here, we describe that cell lines with acquired resistance to vemurafenib show reactivation of MAPK signaling and upregulation of cyclin D1 and are sensitive to inhibition of LY2835219, a selective inhibitor of cyclin-dependent kinase (CDK) 4/6. LY2835219 was demonstrated to inhibit growth of melanoma A375 tumor xenografts and delay tumor recurrence in combination with vemurafenib. Furthermore, we developed an in vivo vemurafenib-resistant model by continuous administration of vemurafenib in A375 xenografts. Consistently, we found that MAPK is reactivated and cyclin D1 is elevated in vemurafenib-resistant tumors, as well as in the resistant cell lines derived from these tumors. Importantly, LY2835219 exhibited tumor growth regression in a vemurafenib-resistant model. Mechanistic analysis revealed that LY2835219 induced apoptotic cell death in a concentration-dependent manner in vemurafenib-resistant cells whereas it primarily mediated cell-cycle G1 arrest in the parental cells. Similarly, RNAi-mediated knockdown of cyclin D1 induced significantly higher rate of apoptosis in the resistant cells than in parental cells, suggesting that elevated cyclin D1 activity is important for the survival of vemurafenib-resistant cells. Altogether, we propose that targeting cyclin D1-CDK4/6 signaling by LY2835219 is an effective strategy to overcome MAPK-mediated resistance to B-RAF inhibitors in B-RAF V600E melanoma.
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Vipin
authors.initials: V
authors.lastname: Yadav
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Teresa F
authors.initials: TF
authors.lastname: Burke
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Huber
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Van Horn
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Youyan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: Sean G
authors.initials: SG
authors.lastname: Buchanan
authors.affiliation: Oncology Business Unit, Eli Lilly and Company, Indianapolis, Indiana.
authors.forename: Edward M
authors.initials: EM
authors.lastname: Chan
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana.
authors.forename: James J
authors.initials: JJ
authors.lastname: Starling
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana. beckmann_richard_p@lilly.com sbpeng@lilly.com.
authors.forename: Richard P
authors.initials: RP
authors.lastname: Beckmann
authors.affiliation: Oncology Discovery Research, Lilly Research Laboratories, Indianapolis, Indiana. beckmann_richard_p@lilly.com sbpeng@lilly.com.
authors.forename: Sheng-Bin
authors.initials: SB
authors.lastname: Peng
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001562:Benzimidazoles / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D019938:Cyclin D1 / Q000378:metabolism*; D051358:Cyclin-Dependent Kinase 4 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051361:Cyclin-Dependent Kinase 6 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D008819:Mice, Nude; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D013449:Sulfonamides / Q000494:pharmacology; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects; D000077484:Vemurafenib; D023041:Xenograft Model Antitumor Assays
chemical_list: D000631:Aminopyridines; D001562:Benzimidazoles; C530104:CCND1 protein, human; D007211:Indoles; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D019938:Cyclin D1; D000077484:Vemurafenib; C000590451:abemaciclib; C495900:CDK4 protein, human; C495911:CDK6 protein, human; D051358:Cyclin-Dependent Kinase 4; D051361:Cyclin-Dependent Kinase 6