Ritonavir reverses resistance to docetaxel and cabazitaxel in prostate cancer cells with acquired resistance to docetaxel

Abstract

Aim: Docetaxel is a microtubule-stabilizing drug used for the treatment of several cancers, including prostate cancer. Resistance to docetaxel can either occur through intrinsic resistance or develop under therapeutic pressure, i.e., acquired resistance. A possible explanation for the occurrence of acquired resistance to docetaxel is increased drug efflux via P-glycoprotein (P-gp) drug transporters.

Methods: We have generated docetaxel-resistant cell lines DU-145DOC10 and 22Rv1DOC8 by exposing parental cell lines DU-145DOC and 22Rv1 to increasing levels of docetaxel. Gene expression levels between DU-145DOC10 and 22Rv1DOC8 were compared with those of their respective originator cell lines. Both parental and resistant cell lines were treated with the taxane drugs docetaxel and cabazitaxel in combination with the P-gp/CYP3A4 inhibitor ritonavir and the P-gp inhibitor elacridar.

Results: In the docetaxel-resistant cell lines DU-145DOC10 and 22Rv1DOC8, the ABCB1 (P-gp) gene was highly up-regulated. Expression of the P-gp protein was also significantly increased in the docetaxel-resistant cell lines in a Western blotting assay. The addition of ritonavir to docetaxel resulted in a return of the sensitivity to docetaxel in the DU-145DOC10 and 22Rv1DOC8 to a level similar to the sensitivity in the originator cells. We found that these docetaxel-resistant cell lines could also be re-sensitized to cabazitaxel in a similar manner. In a Caco-2 P-gp transporter assay, functional inhibition of P-gp-mediated transport of docetaxel with ritonavir was demonstrated.

Conclusion: Our results demonstrate that ritonavir restores sensitivity to both docetaxel and cabazitaxel in docetaxel-resistant cell lines, most likely by inhibiting P-gp-mediated drug efflux.

Keywords

Prostate cancer / docetaxel resistance / cabazitaxel resistance / P-gp / ritonavir

References

[1] Berthold DR,Soban F,Eisenberger M.Docetaxel plus prednisone or mitoxantrone plus prednisone for advanced prostate cancer: updated survival in the TAX 327 study.J Clin Oncol2008;26:242-5

[2] Petrylak DP,Hussain MHA.Docetaxel and estramustine compared with mitoxantrone and prednisone for advanced refractory prostate cancer.N Engl J Med2004;351:1513-20

[3] Waghray D.Inhibit or evade multidrug resistance P-glycoprotein in cancer treatment.J Med Chem2018;61:5108-21 PMCID:PMC6281405

[4] Loos NHC,Schinkel AH.The inhibitory and inducing effects of ritonavir on hepatic and intestinal CYP3A and other drug-handling proteins.Biomed Pharmacother2023;162:114636 PMCID:PMC10065864

[5] Sevrioukova IF.Structure and mechanism of the complex between cytochrome P4503A4 and ritonavir.Proc Natl Acad Sci U S A2010;107:18422-7 PMCID:PMC2973003

[6] Drewe J,Fricker G,Beglinger C.HIV protease inhibitor ritonavir: a more potent inhibitor of P-glycoprotein than the cyclosporine analog SDZ PSC 833.Biochem Pharmacol1999;57:1147-52

[7] Eek D,Mazar I.Patient-reported preferences for oral versus intravenous administration for the treatment of cancer: a review of the literature.Patient Prefer Adherence2016;10:1609-21 PMCID:PMC5003561

[8] van Waterschoot RAB,Wagenaar E.Absence of both cytochrome P450 3A and P-glycoprotein dramatically increases docetaxel oral bioavailability and risk of intestinal toxicity.Cancer Res2009;69:8996-9002

[9] Oostendorp RL,Rosing H.Coadministration of ritonavir strongly enhances the apparent oral bioavailability of docetaxel in patients with solid tumors.Clin Cancer Res2009;15:4228-33

[10] de Weger VA,Koolen SLW.A phase I dose escalation study of once-weekly oral administration of docetaxel as ModraDoc001 capsule or ModraDoc006 tablet in combination with ritonavir.Clin Cancer Res2019;25:5466-74

[11] Vermunt MAC,Devriese LA.ModraDoc006, an oral docetaxel formulation in combination with ritonavir (ModraDoc006/r), in metastatic castration-resistant prostate cancer patients: a phase Ib study.Cancer Rep2021;4:e1367 PMCID:PMC8388171

[12] Ikezoe T,Takeuchi T.HIV-1 protease inhibitor, ritonavir: a potent inhibitor of CYP3A4, enhanced the anticancer effects of docetaxel in androgen-independent prostate cancer cells in vitro and in vivo.Cancer Res2004;64:7426-31

[13] Hendrikx JJMA,Song JY.Ritonavir inhibits intratumoral docetaxel metabolism and enhances docetaxel antitumor activity in an immunocompetent mouse breast cancer model.Int J Cancer2016;138:758-69

[14] van Eijk M, Boosman RJ, Schinkel AH, Huitema ADR, Beijnen JH. Cytochrome P450 3A4, 3A5, and 2C8 expression in breast, prostate, lung, endometrial, and ovarian tumors: relevance for resistance to taxanes.Cancer Chemother Pharmacol2019;84:487-99 PMCID:PMC6682574

[15] Hwang C.Overcoming docetaxel resistance in prostate cancer: a perspective review.Ther Adv Med Oncol2012;4:329-40 PMCID:PMC3481557

[16] Galletti G,Lam L.Mechanisms of resistance to systemic therapy in metastatic castration-resistant prostate cancer.Cancer Treat Rev2017;57:16-27

[17] Maloney SM,Morejon-Lasso LV.Mechanisms of taxane resistance.Cancers2020;12:3323 PMCID:PMC7697134

[18] Gjyrezi A,Voznesensky O.Taxane resistance in prostate cancer is mediated by decreased drug-target engagement.J Clin Invest2020;130:3287-98 PMCID:PMC7259995

[19] Pokharel D,Oenarto V,Bebawy M.Proteins regulating the intercellular transfer and function of P-glycoprotein in multidrug-resistant cancer.Ecancermedicalscience2017;11:768 PMCID:PMC5636210

[20] Kato T,Kameyama K.Serum exosomal P-glycoprotein is a potential marker to diagnose docetaxel resistance and select a taxoid for patients with prostate cancer.Urol Oncol2015;33:385.e15-20

[21] de Bono JS, Oudard S, Ozguroglu M, et al; TROPIC Investigators. Prednisone plus cabazitaxel or mitoxantrone for metastatic castration-resistant prostate cancer progressing after docetaxel treatment: a randomised open-label trial.Lancet2010;376:1147-54

[22] O’Neill AJ,Dowling C.Characterisation and manipulation of docetaxel resistant prostate cancer cell lines.Mol Cancer2011;10:126 PMCID:PMC3203088

[23] Zhu Y,Nadiminty N.Inhibition of ABCB1 expression overcomes acquired docetaxel resistance in prostate cancer.Mol Cancer Ther2013;12:1829-36 PMCID:PMC3947549

[24] Oprea-Lager DE,Van Moorselaar RJA,Hoekstra OS.ABCC4 Decreases docetaxel and not cabazitaxel efficacy in prostate cancer cells in vitro.Anticancer Res2013;33:387-91

[25] Monteverde M,Fischel JL.Combination of docetaxel and vandetanib in docetaxel-sensitive or resistant PC3 cell line.Urol Oncol2013;31:776-86

[26] Zhu S,Li S,Tang Y.Synergistic antitumor activities of docetaxel and octreotide associated with apoptotic-upregulation in castration-resistant prostate cancer.PLoS One2014;9:e91817 PMCID:PMC3954761

[27] Zhu Y,Armstrong C,Sandher A.Antiandrogens inhibit ABCB1 efflux and ATPase activity and reverse docetaxel resistance in advanced prostate cancer.Clin Cancer Res2015;21:4133-42 PMCID:PMC4573793

[28] Lin JZ,De W.Targeting AXL overcomes resistance to docetaxel therapy in advanced prostate cancer.Oncotarget2017;8:41064-77 PMCID:PMC5522277

[29] Luty M,Łabędź-Masłowska A.Fenofibrate augments the sensitivity of drug-resistant prostate cancer cells to docetaxel.Cancers2019;11:77 PMCID:PMC6356694

[30] Wang Y,Chen CZ.Therapeutic targeting of MDR1 expression by RORγ antagonists resensitizes cross-resistant CRPC to taxane via coordinated induction of cell death programs.Mol Cancer Ther2020;19:364-74

[31] Lima TS,Souza LDO.Molecular profiling of docetaxel-resistant prostate cancer cells identifies multiple mechanisms of therapeutic resistance.Cancers2021;13:1290 PMCID:PMC7998254

[32] Ma Y,Peng Z.Identification of mutations, gene expression changes and fusion transcripts by whole transcriptome RNAseq in docetaxel resistant prostate cancer cells.Springerplus2016;5:1861 PMCID:PMC5078122

[33] Marín-Aguilera M,Kalko SG.Identification of docetaxel resistance genes in castration-resistant prostate cancer.Mol Cancer Ther2012;11:329-39

[34] Ahmed SA,Walsh JE.A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: an alternative to [³H]thymidine incorporation assay.J Immunol Methods1994;170:211-24

[35] Berger DP,Winterhalter BR.The clonogenic assay with human tumor xenografts: evaluation, predictive value and application for drug screening.Ann Oncol1990;1:333-41

[36] Law CW,Shi W.voom: Precision weights unlock linear model analysis tools for RNA-seq read counts.Genome Biol2014;15:R29 PMCID:PMC4053721

[37] Cui Y,Schnell D,MacLean C.In-depth characterization of epiintestinal microtissue as a model for intestinal drug absorption and metabolism in human.Pharmaceutics2020;12:405 PMCID:PMC7284918

[38] Szakács G,Ludwig JA,Gottesman MM.Targeting multidrug resistance in cancer.Nat Rev Drug Discov2006;5:219-34

[39] Kumar S,Chamala S.Ritonavir blocks AKT signaling, activates apoptosis and inhibits migration and invasion in ovarian cancer cells.Mol Cancer2009;8:26 PMCID:PMC2691728

[40] Gills JJ,Tsurutani J.Nelfinavir, a lead HIV protease inhibitor, is a broad-spectrum, anticancer agent that induces endoplasmic reticulum stress, autophagy, and apoptosis in vitro and in vivo.Clin Cancer Res2007;13:5183-94

[41] Liu J,Zhu D.Establishment and characterization of a docetaxel-resistant human prostate cancer cell line.Oncol Lett2020;20:230 PMCID:PMC7499980

[42] Mita AC,Mita MM.Cabazitaxel: more than a new taxane for metastatic castrate-resistant prostate cancer?.Clin Cancer Res2012;18:6574-9

[43] Duran GE,Weitz D.Cabazitaxel is more active than first-generation taxanes in ABCB1(+) cell lines due to its reduced affinity for P-glycoprotein.Cancer Chemother Pharmacol2018;81:1095-103

[44] Clinical pharmacology NDA review. Available from: https://www.fda.gov/media/105653/download. [Last accessed on 31 Jan 2024]

[45] Machioka K,Kadono Y.Establishment and characterization of two cabazitaxel-resistant prostate cancer cell lines.Oncotarget2018;9:16185-96 PMCID:PMC5882326

[46] Lombard AP,Armstrong CM.ABCB1 mediates cabazitaxel-docetaxel cross-resistance in advanced prostate cancer.Mol Cancer Ther2017;16:2257-66 PMCID:PMC5628132

[47] Wils P,Warnery A.Polarized transport of docetaxel and vinblastine mediated by P-glycoprotein in human intestinal epithelial cell monolayers.Biochem Pharmacol1994;48:1528-30

[48] Sekino Y.Molecular mechanisms of docetaxel resistance in prostate cancer.Cancer Drug Resist2020;3:676-85 PMCID:PMC8992564