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]	Malard F, Mohty M. Acute lymphoblastic leukaemia. Lancet, 2020,395(10230):1146–1162
[2]	Graux C. Biology of acute lymphoblastic leukemia (ALL): clinical and therapeutic relevance. Transfus Apher Sci, 2011,44(2):183–189
[3]	Leonard J, Stock W. Progress in adult ALL: incorporation of new agents to frontline treatment. Hematology Am Soc Hematol Educ Program, 2017,2017(1):28–36
[4]	Ribera JM, Garcia O, Gil C, 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]	DeAngelo DJ, Stevenson KE, Dahlberg SE, 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]	Huang W, Sun X, Li Y, 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]	Feng L, Chen K, Huang W, et al. Pharmacological targeting PIKfyve and tubulin as an effective treatment strategy for double-hit lymphoma. Cell Death Discov, 2022,8(1):39
[8]	Sbrissa D, Ikonomov OC, Shisheva A. Phosphatidylinositol 3-phosphate-interacting domains in PIKfyve. Binding specificity and role in PIKfyve. Endomenbrane localization. J Biol Chem, 2002,277(8):6073–6079
[9]	Stenmark H, Aasland R, Toh BH, 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]	Kim SM, Roy SG, Chen B, et al. Targeting cancer metabolism by simultaneously disrupting parallel nutrient access pathways. J Clin Invest, 2016,126(11):4088–4102
[11]	Krishna S, Palm W, Lee Y, et al. PIKfyve Regulates Vacuole Maturation and Nutrient Recovery following Engulfment. Dev Cell, 2016,38(5):536–547
[12]	Gayle S, Landrette S, Beeharry N, 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]	Janke C, Magiera MM. The tubulin code and its role in controlling microtubule properties and functions. Nat Rev Mol Cell Biol, 2020,21(6):307–326
[14]	Wloga D, Joachimiak E, Fabczak H. Tubulin Post-Translational Modifications and Microtubule Dynamics. Int J Mol Sci, 2017,18(10):2207
[15]	Florian S, Mitchison TJ. Anti-Microtubule Drugs. Methods Mol Biol, 2016,1413:403–421
[16]	Maltese WA, Overmeyer JH. Methuosis: nonapoptotic cell death associated with vacuolization of macropinosome and endosome compartments. Am J Pathol, 2014,184(6):1630–1642
[17]	Overmeyer JH, Young AM, Bhanot H, 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]	Sorger PK, Dobles M, Tournebize R, et al. Coupling cell division and cell death to microtubule dynamics. Curr Opin Cell Biol, 1997,9(6):807–814
[19]	Horio T, Murata T. The role of dynamic instability in microtubule organization. Front Plant Sci, 2014,5:511
[20]	Vandenabeele P, Galluzzi L, Vanden Berghe T, et al. Molecular mechanisms of necroptosis: an ordered cellular explosion. Nat Rev Mol Cell Biol, 2010,11(10):700–714
[21]	Visvader JE. Cells of origin in cancer. Nature, 2011,469(7330):314–322
[22]	Liu JS, Huo CY, Cao HH, 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]	Wang BJ, Zheng WL, Feng NN, 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]	Li H, Dan C, Gong X, 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]	Dolcet X, Llobet D, Pallares J, et al. NF-kB in development and progression of human cancer. Virchows Arch, 2005,446(5):475–482
[26]	Haupt Y, Rowan S, Shaulian E, et al. p53 mediated apoptosis in HeLa cells: transcription dependent and independent mechanisms. Leukemia, 1997,11 Suppl 3:337–339
[27]	Thompson EB. The many roles of c-Myc in apoptosis. Annu Rev Physiol, 1998,60:575–600
[28]	McMahon SB. MYC and the control of apoptosis. Cold Spring Harb Perspect Med, 2014,4(7):a014407
[29]	Asano T, Yao Y, Zhu J, 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]	Koul D, Yao Y, Abbruzzese JL, 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]	Fujiwara Y, Hosokawa Y, Watanabe K, 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]	Onishi K, Higuchi M, Asakura T, et al. The PI3K-Akt pathway promotes microtubule stabilization in migrating fibroblasts. Genes Cells, 2007,12(4):535–546