Abstract
Nuphar alkaloids, originally isolated from water lilies, induce apoptosis in mammalian cells in less than 1 h, making them possibly the fastest known inducers. However, the mechanism by which this rapid apoptosis occurs remains unknown. We have investigated canonical aspects of apoptosis to determine how the nuphar alkaloid, (+)-6-hydroxythiobinupharidine (6HTBN), induces apoptosis. 6HTBN induced rapid apoptosis in various leukemia, lymphoma, and carcinoma cell lines, suggesting that its mechanism is cell-type independent. It also circumvented resistance of patient-derived chronic lymphocytic leukemia cells generated by co-culture on survival-promoting stroma. Intriguingly, 6HTBN failed to induce apoptosis in platelets. The mechanism of apoptosis involves activation of caspase 9 and caspase 3, but not caspase 8 as previously reported. The release of cytochrome c from mitochondria occurred even in the absence of BAX/BAK and in cells that retained mitochondrial membrane potential. These results suggest a novel mechanism of apoptosis that has previously not been reported. The molecular target of the nuphar alkaloids remains to be determined.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Achmatowicz O, Bellen Z. Alkaloids of Nuphar luteum (L) sm. Isolation of alkaloids containing sulphur. Tetrahedron Lett. 1962;3:1121–4.
Bates DJP, Salerni BL, Lowrey CH, Eastman A. Vinblastine sensitizes leukemia cells to cyclin-dependent kinase inhibitors inducing acute cell cycle phase-independent apoptosis. Cancer Biol Therap. 2011;12:314–25.
Ganju N, Eastman A. Zinc inhibits Bax and Bak activation and cytochrome c release induced by chemical inducers of apoptosis but not by death-receptor-initiated pathways. Cell Death Differ. 2003;10:652–61.
Han J, Goldstein LA, Gastman BR, Rabinowitz ZA, Wang G-Q, Fang B, et al. Differential involvement of Bax and Bak in TRAIL-mediated apoptosis of leukemic T cells. Leukemia. 2004;18:16711680.
Jansen DJ, Shenvi RA. Synthesis of (−)-neothiobinupharidine. J Am Chem Soc. 2013;135:1209–12.
Korotkov A, Li H, Chapman C, Povinelli L, MacMillan J, Eastman A, et al. Total synthesis and biological evaluation of both enantiomers of several hydroxylated dimeric nuphar alkaloids. Angew Chem Int Ed Engl. 2015;54:10604–7.
Lacharity JJ, Fournier J, Lu P, Mailyan AK, Herrmann AT, Zakarian A. Total synthesis of unsymmetrically oxidized nuphar thioalkaloids via copper-catalyzed thiolane assembly. J Am Chem Soc. 2017;139:13272–5.
Leverson JD, Zhang H, Chen J, Tahir SK, Phillips DC, Xue J, et al. Potent and selective small-molecule MCL-1 inhibitors demonstrate on-target cancer cell killing activity as single agents and in combination with ABT-263 (navitoclax). Cell Death Dis. 2015;6:e1590.
Li H, Korotkov A, Chapman CW, Eastman A, Wu J. Enantioselective formal synthesis of 11 nuphar alkaloids and discovery of the first known apoptotic monomeric analogs. Angew Chem Int Ed Engl. 2016;55:3509–13.
Li H, Cooke TJ, Korotkov A, Chapman CW, Eastman A. Wu J. Stereoselective synthesis and biological evaluation of C1-epimeric and desmethyl monomeric nuphar analogues. J Org Chem. 2017;82:2648–55.
Mallick DJ, Soderquist RS, Bates D, Eastman A. Confounding off-target effects of BH3 mimetics at commonly used concentrations: MIM1, UMI77 and A1210477. Cell Death Dis. 2019;10:185.
Matsuda H, Yoshida K, Miyagawa K, Nemoto Y, Asao Y, Yoskikawa M. Nuphar alkaloids with immediately apoptosis-inducing activity from Nuphar pumilum and their structural requirements for activity. Bioorg Med Chem Lett. 2006;16:1567–73.
Neron S, Pelletier A, Chevrier MC, Monier G, Lemieux R, Darveau A. Induction of LFA-1 independent human B cell proliferation and differentiation by binding of CD40 with its ligand. Immunol Investig. 1996;25:79–89.
Ozer J, Eisner N, Ostrozhenkova E, Bacher A, Eisenreich W, Benharroch D, et al. Nuphar lutea thioalkaloids inhibit the nuclear factor kappa B pathway, potentiate apoptosis and are synergistic with cisplatin and etoposide. Cancer Biol Ther. 2009;8:1860–8.
Perelman A, Wachtel C, Cohen M, Haupt S, Shapiro H, Tzur A. JC-1: alternative excitation wavelengths facilitate mitochondrial membrane potential cytometry. Cell Death Dis. 2012;3:e430.
Single B, Leist M, Nicotera P. Simultaneous release of adenylate kinase and cytochrome c in cell death. Cell Death Differ. 1998;5:1001–3.
Soderquist RS, Bates DJP, Danilov AV, Eastman A. Gossypol overcomes stroma-mediated resistance to the BCL-2 inhibitor ABT-737 in chronic lymphocytic leukemia. Leukemia. 2013;27:2262–4.
Soderquist RS, Danilov AV, Eastman A. Gossypol increases expression of the pro-apoptotic protein through a novel mechanism involving phospholipase A2, cytoplasmic calcium and endoplasmic reticulum stress. J Biol Chem. 2014;289:16190–9.
Tada N, Jansen DJ, Mower MP, Blewett MM, Umotoy JC, Cravatt BF, et al. Synthesis and sulfur electrophilicity of the Nuphar thiaspirane pharmacophore. ACS Cent Sci. 2016;2:401–8.
Vander Heiden MG, Chandel NS, Williamson EK, Thompson CB. Bcl-xL regulates the membrane potential and volume homeostasis of mitochondria. Cell. 1997;91:627–37.
Zhang H, Nimmer PM, Tahir SK, Chen J, Fryer RM, Hahn KR, et al. Bcl-2 family proteins are essential for platelet survival. Cell Death Differ. 2017;14:943–51.
Funding
This research was supported by grant RSG-13-011-01-CDD from the American Cancer Society to JW and AE, a pilot grant from the COBRE Institute for Biomolecular Targeting (P20GM113132), and an NCI Cancer Center Support Grant 5P30 CA023108 to the Norris Cotton Cancer Center.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mallick, D.J., Korotkov, A., Li, H. et al. Nuphar alkaloids induce very rapid apoptosis through a novel caspase-dependent but BAX/BAK-independent pathway. Cell Biol Toxicol 35, 435–443 (2019). https://doi.org/10.1007/s10565-019-09469-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10565-019-09469-5