Pan‑RAF inhibitor LY3009120 is highly synergistic with low‑dose cytarabine, but not azacitidine, in acute myeloid leukemia with <em>RAS</em> mutations

Park,Jihyun; Park,Hyejoo; Byun,Ja Min; Hong,Junshik; Shin,Dong-Yeop; Koh,Youngil; Yoon,Sung-Soo

doi:10.3892/ol.2021.13006

Pan‑RAF inhibitor LY3009120 is highly synergistic with low‑dose cytarabine, but not azacitidine, in acute myeloid leukemia with RAS mutations

Authors:
- Jihyun Park
- Hyejoo Park
- Ja Min Byun
- Junshik Hong
- Dong-Yeop Shin
- Youngil Koh
- Sung-Soo Yoon
View Affiliations

Affiliations: Cancer Research Institute, Seoul National University College of Medicine, Seoul 03080, Republic of Korea, Department of Internal Medicine, Division of Hematology and Medical Oncology, Seoul National University Hospital, Seoul 03080, Republic of Korea, Hematology Oncology Department, Center for Medical Innovation, Seoul National University Hospital, Seoul 03082, Republic of Korea
Published online on: August 20, 2021 https://doi.org/10.3892/ol.2021.13006
Article Number: 745

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Alterations in RAS oncogenes have been implicated in various types of cancer, including acute myeloid leukemia (AML). Considering that currently, there are no targeted therapies for patients with RAS‑mutated AML despite the poor outcomes, RAF may be a potential target for AML. In this study, we first analyzed the efficacy of different MAPK inhibitors in AML cell lines. We found that LY3009120, a pan‑RAF inhibitor, significantly decreased cell survival in RAS‑mutated AML cell lines. We then investigated the synergistic effects of LY3009120 with either cytarabine or azacitidine. We found that the combination of low‑dose cytarabine and LY3009120 showed a synergistic effect in NRAS‑mutated HL‑60 cells and KRAS‑mutated NB4 cells. This effect was caused by a decrease in proliferation, induction of apoptosis, and cell growth arrest through a decrease in phosphorylated MEK and ERK along with a cytotoxic response occurring specifically for the RAS mutation of the pan‑RAF inhibitor LY3009120. In addition, we confirmed that combination treatment with low‑dose cytarabine and LY3009120 led to an increase in apoptosis in primary AML cells. Our findings indicate that combination therapy with pan‑RAF inhibitor LY3009120 and low‑dose cytarabine may be a promising treatment strategy for RAS‑mutated AML.

View Figures

View References

1	Almeida AM and Ramos F: Acute myeloid leukemia in the older adults. Leuk Res Rep. 6:1–7. 2016.PubMed/NCBI
2	Papaemmanuil E, Gerstung M, Bullinger L, Gaidzik VI, Paschka P, Roberts ND, Potter NE, Heuser M, Thol F, Bolli N, et al: Genomic classification and prognosis in acute myeloid leukemia. N Engl J Med. 374:2209–2221. 2016. View Article : Google Scholar : PubMed/NCBI
3	Lagunas-Rangel FA, Chavez-Valencia V, Gomez-Guijosa MA and Cortes-Penagos C: Acute myeloid leukemia-genetic alterations and their clinical prognosis. Int J Hematol Oncol Stem Cell Res. 11:328–339. 2017.PubMed/NCBI
4	Short NJ, Rytting ME and Cortes JE: Acute myeloid leukaemia. Lancet. 392:593–606. 2018. View Article : Google Scholar : PubMed/NCBI
5	Hobbs GA, Der CJ and Rossman KL: RAS isoforms and mutations in cancer at a glance. J Cell Sci. 129:1287–1292. 2016.PubMed/NCBI
6	Ward AF, Braun BS and Shannon KM: Targeting oncogenic Ras signaling in hematologic malignancies. Blood. 120:3397–3406. 2012. View Article : Google Scholar : PubMed/NCBI
7	Kadia TM, Kantarjian H, Kornblau S, Borthakur G, Faderl S, Freireich EJ, Luthra R, Garcia-Manero G, Pierce S, Cortes J and Ravandi F: Clinical and proteomic characterization of acute myeloid leukemia with mutated RAS. Cancer. 118:5550–5559. 2012. View Article : Google Scholar : PubMed/NCBI
8	Steelman LS, Franklin RA, Abrams SL, Chappell W, Kempf CR, Bäsecke J, Stivala F, Donia M, Fagone P, Nicoletti F, et al: Roles of the Ras/Raf/MEK/ERK pathway in leukemia therapy. Leukemia. 25:1080–1094. 2011. View Article : Google Scholar : PubMed/NCBI
9	Downward J: Targeting RAS signalling pathways in cancer therapy. Nat Rev Cancer. 3:11–22. 2003. View Article : Google Scholar : PubMed/NCBI
10	Kornblau SM, Womble M, Qiu YH, Jackson CE, Chen W, Konopleva M, Estey EH and Andreeff M: Simultaneous activation of multiple signal transduction pathways confers poor prognosis in acute myelogenous leukemia. Blood. 108:2358–2365. 2006. View Article : Google Scholar : PubMed/NCBI
11	Steelman LS, Abrams SL, Whelan J, Bertrand FE, Ludwig DE, Bäsecke J, Libra M, Stivala F, Milella M, Tafuri A, et al: Contributions of the Raf/MEK/ERK, PI3K/PTEN/Akt/mTOR and Jak/STAT pathways to leukemia. Leukemia. 22:686–707. 2008. View Article : Google Scholar : PubMed/NCBI
12	Cox AD, Fesik SW, Kimmelman AC, Luo J and Der CJ: Drugging the undruggable RAS: Mission possible? Nat Rev Drug Discov. 13:828–851. 2014. View Article : Google Scholar : PubMed/NCBI
13	Coombs CC, Tallman MS and Levine RL: Molecular therapy for acute myeloid leukaemia. Nat Rev Clin Oncol. 13:305–318. 2016. View Article : Google Scholar : PubMed/NCBI
14	Kavanagh S, Murphy T, Law A, Yehudai D, Ho JM, Chan S and Schimmer AD: Emerging therapies for acute myeloid leukemia: Translating biology into the clinic. JCI Insight. 2:e956792017. View Article : Google Scholar : PubMed/NCBI
15	Lu S, Jang H, Gu S, Zhang J and Nussinov R: Drugging Ras GTPase: A comprehensive mechanistic and signaling structural view. Chem Soc Rev. 45:4929–4952. 2016. View Article : Google Scholar : PubMed/NCBI
16	Schmieder R, Puehler F, Neuhaus R, Kissel M, Adjei AA, Miner JN, Mumberg D, Ziegelbauer K and Scholz A: Allosteric MEK1/2 inhibitor refametinib (BAY 86-9766) in combination with sorafenib exhibits antitumor activity in preclinical murine and rat models of hepatocellular carcinoma. Neoplasia. 15:1161–1171. 2013. View Article : Google Scholar : PubMed/NCBI
17	Dickson MA, Gordon MS, Edelman G, Bendell JC, Kudchadkar RR, LoRusso PM, Johnston SH, Clary DO and Schwartz GK: Phase I study of XL281 (BMS-908662), a potent oral RAF kinase inhibitor, in patients with advanced solid tumors. Invest New Drugs. 33:349–356. 2015. View Article : Google Scholar : PubMed/NCBI
18	Mendzelevski B, Ferber G, Janku F, Li BT, Sullivan RJ, Welsch D, Chi W, Jackson J, Weng O and Sager PT: Effect of ulixertinib, a novel ERK1/2 inhibitor, on the QT/QTc interval in patients with advanced solid tumor malignancies. Cancer Chemother Pharmacol. 81:1129–1141. 2018. View Article : Google Scholar : PubMed/NCBI
19	Ryan MB and Corcoran RB: Therapeutic strategies to target RAS-mutant cancers. Nat Rev Clin Oncol. 15:709–720. 2018. View Article : Google Scholar : PubMed/NCBI
20	Borthakur G, Popplewell L, Boyiadzis M, Foran J, Platzbecker U, Vey N, Walter RB, Olin R, Raza A, Giagounidis A, et al: Activity of the oral mitogen-activated protein kinase kinase inhibitor trametinib in RAS-mutant relapsed or refractory myeloid malignancies. Cancer. 122:1871–1879. 2016. View Article : Google Scholar : PubMed/NCBI
21	Jain N, Curran E, Iyengar NM, Diaz-Flores E, Kunnavakkam R, Popplewell L, Kirschbaum MH, Karrison T, Erba HP, Green M, et al: Phase II study of the oral MEK inhibitor selumetinib in advanced acute myelogenous leukemia: A University of Chicago phase II consortium trial. Clin Cancer Res. 20:490–498. 2014. View Article : Google Scholar : PubMed/NCBI
22	Maiti A, Naqvi K, Kadia TM, Borthakur G, Takahashi K, Bose P, Daver NG, Patel A, Alvarado Y, Ohanian M, et al: Phase II trial of MEK inhibitor binimetinib (MEK162) in RAS-mutant acute myeloid leukemia. Clin Lymphoma Myeloma Leuk. 19:142–148. 2019. View Article : Google Scholar : PubMed/NCBI
23	Peng SB, Henry JR, Kaufman MD, Lu WP, Smith BD, Vogeti S, Rutkoski TJ, Wise S, Chun L, Zhang Y, et al: Inhibition of RAF isoforms and active dimers by LY3009120 leads to anti-tumor activities in RAS or BRAF mutant cancers. Cancer Cell. 28:384–398. 2015. View Article : Google Scholar : PubMed/NCBI
24	Keum H, Kim TW, Kim Y, Seo C, Son Y, Kim J, Kim D, Jung W, Whang CH and Jon S: Bilirubin nanomedicine alleviates psoriatic skin inflammation by reducing oxidative stress and suppressing pathogenic signaling. J Control Release. 325:359–369. 2020. View Article : Google Scholar : PubMed/NCBI
25	Lee SH, Kim DK, Seo YR, Woo KM, Kim CS and Cho MH: Nickel (II)-induced apoptosis and G2/M enrichment. Exp Mol Med. 30:171–176. 1998. View Article : Google Scholar : PubMed/NCBI
26	Wang HM, Chen CY and Wu PF: Isophilippinolide A arrests cell cycle progression and induces apoptosis for anticancer inhibitory agents in human melanoma cells. J Agric Food Chem. 62:1057–1065. 2014. View Article : Google Scholar : PubMed/NCBI
27	Chou TC: Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 70:440–446. 2010. View Article : Google Scholar : PubMed/NCBI
28	Zhao L, Au JL and Wientjes MG: Comparison of methods for evaluating drug-drug interaction. Front Biosci (Elite Ed). 2:241–249. 2010.PubMed/NCBI
29	Kufe DW, Griffin JD and Spriggs DR: Cellular and clinical pharmacology of low-dose ara-C. Semin Oncol. 12 (Suppl 3):S200–S207. 1985.
30	Spriggs D, Griffin J, Wisch J and Kufe D: Clinical pharmacology of low-dose cytosine arabinoside. Blood. 65:1087–1089. 1985. View Article : Google Scholar : PubMed/NCBI
31	Chen L, Guo P, Zhang Y, Li X, Jia P, Tong J and Li J: Autophagy is an important event for low-dose cytarabine treatment in acute myeloid leukemia cells. Leuk Res. 60:44–52. 2017. View Article : Google Scholar : PubMed/NCBI
32	Liu Z, Chen J, Wang H, Ding K, Li Y, de Silva A, Sehgal V, Burbano JL, Sundararaj R, Gamage J, et al: Chidamide shows synergistic cytotoxicity with cytarabine via inducing G0/G1 arrest and apoptosis in myelodysplastic syndromes. Am J Transl Res. 9:5631–5642. 2017.PubMed/NCBI
33	Vakana E, Pratt S, Blosser W, Dowless M, Simpson N, Yuan XJ, Jaken S, Manro J, Stephens J, Zhang Y, et al: LY3009120, a panRAF inhibitor, has significant anti-tumor activity in BRAF and KRAS mutant preclinical models of colorectal cancer. Oncotarget. 8:9251–9266. 2017. View Article : Google Scholar : PubMed/NCBI
34	Tambe M, Karjalainen E, Vähä-Koskela M, Bulanova D, Gjertsen BT, Kontro M, Porkka K, Heckman CA and Wennerberg K: Pan-RAF inhibition induces apoptosis in acute myeloid leukemia cells and synergizes with BCL2 inhibition. Leukemia. 34:3186–3196. 2020. View Article : Google Scholar : PubMed/NCBI
35	Hall-Jackson CA, Eyers PA, Cohen P, Goedert M, Boyle FT, Hewitt N, Plant H and Hedge P: Paradoxical activation of Raf by a novel Raf inhibitor. Chem Biol. 6:559–568. 1999. View Article : Google Scholar : PubMed/NCBI
36	Hatzivassiliou G, Song K, Yen I, Brandhuber BJ, Anderson DJ, Alvarado R, Ludlam MJ, Stokoe D, Gloor SL, Vigers G, et al: RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth. Nature. 464:431–435. 2010. View Article : Google Scholar : PubMed/NCBI
37	King AJ, Patrick DR, Batorsky RS, Ho ML, Do HT, Zhang SY, Kumar R, Rusnak DW, Takle AK, Wilson DM, et al: Demonstration of a genetic therapeutic index for tumors expressing oncogenic BRAF by the kinase inhibitor SB-590885. Cancer Res. 66:11100–11105. 2006. View Article : Google Scholar : PubMed/NCBI
38	Wei AH, Strickland SA Jr, Hou JZ, Fiedler W, Lin TL, Walter RB, Enjeti A, Tiong IS, Savona M, Lee S, et al: Venetoclax combined with low-dose cytarabine for previously untreated patients with acute myeloid leukemia: Results from a phase Ib/II study. J Clin Oncol. 37:1277–1284. 2019. View Article : Google Scholar : PubMed/NCBI