Neratinib inhibits proliferation and promotes apoptosis of acute myeloid leukemia cells by activating autophagy‐dependent ferroptosis

Abstract

Acute myeloid leukemia (AML) is a hematologic malignancy with increased lethality.We focused on elucidating the role of Neratinib, a tyrosine kinase inhibitor, in the progression of AML and identify the potential mechanisms. Upon the treatment of Neratinib, autophagy suppressor 3-methyladenine (3-MA) and ferroptosis stimulator Erastin, the viability and proliferation of HL-60 cells were evaluated by cell counting kit-8 and 5-Ethynyl-20-Deoxyuridine staining assays. A flow cytometer was to observe cell cycle and apoptosis. Production of reactive oxygen species (ROS) was tested via 2,7-dichlorodihydrofluorescein diacetate assay. Additionally, malondialdehyde (MDA) content and Fe 2+ activity were examined with commercial kits. LC3-II expression was examined by using immunofluoresence staining. Western blot analysis ascertained the expression of proliferation, apoptosis, ferroptosis and autophagy-associated proteins. It was noted that Neratinib notably mitigated cell viability and proliferation, cut down Ki67 and proliferating cell nuclear antigen expression. Moreover, Neratinib hindered cell cycle at G0/G1 phase whereas exacerbated apoptosis. ROS, MDA and Fe 2+ activities were elevated by Neratinib, coupled with the reduced glutathione peroxidase 4, ferritin heavy chain 1 expression and enhanced acyl-CoA synthetase long-chain family member 4 expression. Furthermore, Neratinib promoted autophagy of HL-60 cells, evidenced by raised LC3-II, ATG5, Beclin1 expression and lessened p62 expression. Importantly, 3-MA eased the impacts of Neratinib on cell ferroptosis, proliferation and apoptosis, which were offset by further administration of Erastin. To conclude, Neratinib could suppress proliferation and promote apoptosis of HL-60 cells through autophagy-dependent ferroptosis.

Targeting ferroptosis for leukemia therapy: exploring novel strategies from its mechanisms and role in leukemia based on nanotechnology

“…Neratinib, a tyrosine kinase inhibitor approved by the U.S. Food and Drug Administration in 2017 for breast cancer treatment, has been found to trigger autophagy-dependent ferroptosis, G0/G1 arrest, and apoptosis in HL-60 cells. A phase 1/2 clinical trial of neratinib is currently underway in pediatric patients with relapsed or refractory cancer, including leukemia (NCT02932280) [ 219 ].…”

Crosstalk between autophagy and metabolism: implications for cell survival in acute myeloid leukemia

“… Methods Drug used Anti-AML effect in pre-clinic research Ref. Autophagy inhibition 3-methyladenine Increase the sensitivity of AML cells to cytarabine both in vitro and in vivo [ 24 ] Chloroquine Enhances the toxicity of AraC to AML cells but appears to be ineffective for patients who have relapsed and for AraC-resistant AML cells [ 3 ] Bafilomycin Potentiates the cytotoxic effects of ATO/ATRA in AML cell lines KG-1 and HL-60 [ 25 ] Wortmannin Impairs expression of anti-apoptotic Bcl-2 family members Bcl-2 and BclXL in primary AML cells via blockade of PI3K/AKT signaling [ 26 ] Autophagy induction Metformin Inhibits Acute Myeloid Leukemia Cell Growth through the AMPK/mTOR Pathway and Autophagic Regulation [ 27 ] Resveratrol Modulates autophagy and induces apoptosis in HL-60 cells [ 28 ] Quercetin Induces autophagy-associated death in HL-60 cells through CaMKKβ/AMPK/mTOR signal pathway [ 29 ] Neratinib Suppresses proliferation and promotes apoptosis of HL-60 cells through autophagy-dependent ferroptosis [ 30 ] ATRA Promotes differentiation in APL cells via autophagy induction [ 31 ] ATPR Triggers ferroptosis and promotes AML differentiation through the regulation of autophagy via iron homeostasis [ 32 ] APL acute promyelocytic leukemia, ATO arsenic trioxide, ATPR 4-Amino-2-Trifluoromethyl-Phenyl Retinate, ATRA all-trans retinoic acid, AraC cytarabine. …”

Molecular Mechanisms of Ferroptosis and Updates of Ferroptosis Studies in Cancers and Leukemia

“…The tyrosine kinase inhibitor Neratinib was approved by the U.S. Food and Drug Administration in 2017 for the treatment of breast cancer, and a phase 1/2 clinical trial of this drug in pediatric patients with relapsed or refractory cancer including leukemia is ongoing (NCT02932280). Recently, neratinib has been shown to induce autophagy-dependent ferroptosis as well as G0/G1 arrest and apoptosis in HL-60 cells [ 193 ].…”