Novel PIKfyve/Tubulin Dual-target Inhibitor as a Promising Therapeutic Strategy for B-cell Acute Lymphoblastic Leukemia

Abstract

Objective

In B-cell acute lymphoblastic leukemia (B-ALL), current intensive chemotherapies for adult patients fail to achieve durable responses in more than 50% of cases, underscoring the urgent need for new therapeutic regimens for this patient population. The present study aimed to determine whether HZX-02-059, a novel dual-target inhibitor targeting both phosphatidylinositol-3-phosphate 5-kinase (PIKfyve) and tubulin, is lethal to B-ALL cells and is a potential therapeutic for B-ALL patients.

Methods

Cell proliferation, vacuolization, apoptosis, cell cycle, and in-vivo tumor growth were evaluated. In addition, Genome-wide RNA-sequencing studies were conducted to elucidate the mechanisms of action underlying the anti-leukemia activity of HZX-02-059 in B-ALL.

Results

HZX-02-059 was found to inhibit cell proliferation, induce vacuolization, promote apoptosis, block the cell cycle, and reduce in-vivo tumor growth. Downregulation of the p53 pathway and suppression of the phosphoinositide 3-kinase (PI3K)/AKT pathway and the downstream transcription factors c-Myc and NF-κB were responsible for these observations.

Conclusion

Overall, these findings suggest that HZX-02-059 is a promising agent for the treatment of B-ALL patients resistant to conventional therapies.

Keywords

B-cell acute lymphoblastic leukemia / dual-target inhibitor / NF-κB / c-Myc / PI3K/AKT / p53

References

[1] MalardF, MohtyM. Acute lymphoblastic leukaemia. Lancet, 2020, 395(10230): 1146-1162

[2] GrauxC. Biology of acute lymphoblastic leukemia (ALL): clinical and therapeutic relevance. Transfus Apher Sci, 2011, 44(2): 183-189

[3] LeonardJ, StockW. Progress in adult ALL: incorporation of new agents to frontline treatment. Hematology Am Soc Hematol Educ Program, 2017, 2017(1): 28-36

[4] RiberaJM, GarciaO, GilC, et al.. Comparison of intensive, pediatric-inspired therapy with nonintensive therapy in older adults aged 55–65 years with Philadelphia chromosome-negative acute lymphoblastic leukemia. Leuk Res, 2018, 68: 79-84

[5] DeAngeloDJ, StevensonKE, DahlbergSE, et al.. Long-term outcome of a pediatric-inspired regimen used for adults aged 18–50 years with newly diagnosed acute lymphoblastic leukemia. Leukemia, 2015, 29(3): 526-534

[6] HuangW, SunX, LiY, et al.. Discovery and Identification of Small Molecules as Methuosis Inducers with in Vivo Antitumor Activities. J Med Chem, 2018, 61(12): 5424-5434

[7] FengL, ChenK, HuangW, et al.. Pharmacological targeting PIKfyve and tubulin as an effective treatment strategy for double-hit lymphoma. Cell Death Discov, 2022, 8(1): 39

[8] SbrissaD, IkonomovOC, ShishevaA. Phosphatidylinositol 3-phosphate-interacting domains in PIKfyve. Binding specificity and role in PIKfyve. Endomenbrane localization. J Biol Chem, 2002, 277(8): 6073-6079

[9] StenmarkH, AaslandR, TohBH, et al.. Endosomal localization of the autoantigen EEA1 is mediated by a zinc-binding FYVE finger. J Biol Chem, 1996, 271(39): 24048-24054

[10] KimSM, RoySG, ChenB, et al.. Targeting cancer metabolism by simultaneously disrupting parallel nutrient access pathways. J Clin Invest, 2016, 126(11): 4088-4102

[11] KrishnaS, PalmW, LeeY, et al.. PIKfyve Regulates Vacuole Maturation and Nutrient Recovery following Engulfment. Dev Cell, 2016, 38(5): 536-547

[12] GayleS, LandretteS, BeeharryN, et al.. Identification of apilimod as a first-in-class PIKfyve kinase inhibitor for treatment of B-cell non-Hodgkin lymphoma. Blood, 2017, 129(13): 1768-1778

[13] JankeC, MagieraMM. The tubulin code and its role in controlling microtubule properties and functions. Nat Rev Mol Cell Biol, 2020, 21(6): 307-326

[14] WlogaD, JoachimiakE, FabczakH. Tubulin Post-Translational Modifications and Microtubule Dynamics. Int J Mol Sci, 2017, 18(10): 2207

[15] FlorianS, MitchisonTJ. Anti-Microtubule Drugs. Methods Mol Biol, 2016, 1413: 403-421

[16] MalteseWA, OvermeyerJH. Methuosis: nonapoptotic cell death associated with vacuolization of macropinosome and endosome compartments. Am J Pathol, 2014, 184(6): 1630-1642

[17] OvermeyerJH, YoungAM, BhanotH, et al.. A chalcone-related small molecule that induces methuosis, a novel form of non-apoptotic cell death, in glioblastoma cells. Mol Cancer, 2011, 10: 69

[18] SorgerPK, DoblesM, TournebizeR, et al.. Coupling cell division and cell death to microtubule dynamics. Curr Opin Cell Biol, 1997, 9(6): 807-814

[19] HorioT, MurataT. The role of dynamic instability in microtubule organization. Front Plant Sci, 2014, 5: 511

[20] VandenabeeleP, GalluzziL, Vanden BergheT, et al.. Molecular mechanisms of necroptosis: an ordered cellular explosion. Nat Rev Mol Cell Biol, 2010, 11(10): 700-714

[21] VisvaderJE. Cells of origin in cancer. Nature, 2011, 469(7330): 314-322

[22] LiuJS, HuoCY, CaoHH, et al.. Aloperine induces apoptosis and G2/M cell cycle arrest in hepatocellular carcinoma cells through the PI3K/Akt signaling pathway. Phytomedicine, 2019, 61: 152843

[23] WangBJ, ZhengWL, FengNN, et al.. The Effects of Autophagy and PI3K/AKT/m-TOR Signaling Pathway on the Cell-Cycle Arrest of Rats Primary Sertoli Cells Induced by Zearalenone. Toxins (Basel), 2018, 10(10): 398

[24] LiH, DanC, GongX, et al.. Sorghumol triterpene inhibits the growth of circulating renal cancer cells by promoting cell apoptosis, G2/M cell cycle arrest and downregulating m-TOR/PI3K/AKT signalling pathway. J BUON, 2019, 24(1): 310-314

[25] DolcetX, LlobetD, PallaresJ, et al.. NF-kB in development and progression of human cancer. Virchows Arch, 2005, 446(5): 475-482

[26] HauptY, RowanS, ShaulianE, et al.. p53 mediated apoptosis in HeLa cells: transcription dependent and independent mechanisms. Leukemia, 1997, 11(Suppl 3): 337-339

[27] ThompsonEB. The many roles of c-Myc in apoptosis. Annu Rev Physiol, 1998, 60: 575-600

[28] McMahonSB. MYC and the control of apoptosis. Cold Spring Harb Perspect Med, 2014, 4(7): a014407

[29] AsanoT, YaoY, ZhuJ, et al.. The PI 3-kinase/Akt signaling pathway is activated due to aberrant Pten expression and targets transcription factors NF-kappaB and c-Myc in pancreatic cancer cells. Oncogene, 2004, 23(53): 8571-8580

[30] KoulD, YaoY, AbbruzzeseJL, et al.. Tumor suppressor MMAC/PTEN inhibits cytokine-induced NFkappaB activation without interfering with the IkappaB degradation pathway. J Biol Chem, 2001, 276(14): 11402-11408

[31] FujiwaraY, HosokawaY, WatanabeK, et al.. Blockade of the phosphatidylinositol-3-kinase-Akt signaling pathway enhances the induction of apoptosis by microtubule-destabilizing agents in tumor cells in which the pathway is constitutively activated. Mol Cancer Ther, 2007, 6(3): 1133-1142

[32] OnishiK, HiguchiM, AsakuraT, et al.. The PI3K-Akt pathway promotes microtubule stabilization in migrating fibroblasts. Genes Cells, 2007, 12(4): 535-546