Abstract
Osteosarcoma has been reported with treatment failure in up to 40% of cases. Our laboratory had identified genes involved in the PPARγ pathway to be associated with doxorubicin (DOX) resistance. We hence used PPARγ agonist pioglitazone (PIO) to modulate DOX resistance. DOX-resistant cell line (143B-DOX) was developed by gradient exposure to DOX. The cytotoxicity to PIO and in combination with DOX was assayed in vitro, followed by HPLC to estimate the metabolites of PIO in the presence of microsomes (HLMs). Gene expression studies revealed the mechanism behind the cytotoxicity of PIO. Further, the effects were evaluated in mice bearing 143B-DOX tumors treated either with PIO (20 mg/kg/p.o or 40 mg/kg/p.o Q1D) alone or in combination with DOX (0.5 mg/kg/i.p Q2W). 143B-DOX was 50-fold resistant over parental cells. While PIO did not show any activity on its own, the addition of HLMs to the cells in culture showed over 80% cell kill within 24 h, possibly due to the metabolites of PIO as determined by HPLC. In combination with DOX, PIO had shown synergistic activity. Additionally, cytotoxicity assay in the presence of HLMs revealed that PIO on its own showed promising activity compared to its metabolites—hydroxy pioglitazone and keto pioglitazone. In vivo studies demonstrated that treatment with 40 mg/kg/p.o PIO alone showed significant activity, followed by a combination with DOX. Gene expression studies revealed that PIO could modulate drug resistance by downregulating MDR1 and IL8. Our study suggests that PIO can modulate DOX resistance in osteosarcoma cells.
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References
Apostoli AJ, Nicol CJB (2012) PPAR medicines and human disease: the ABCs of it all. PPAR Res 2012:1–16. https://doi.org/10.1155/2012/504918
Baba S (2001) Pioglitazone: a review of Japanese clinical studies. Curr Med Res Opin 17:166–189. https://doi.org/10.1185/0300799039117059
Basu-roy U, Han E, Rattanakorn K et al (2016) PPARγ agonists promote differentiation of cancer stem cells by restraining YAP transcriptional activity. Oncotarget 7:60954–60970
Beluzi M, Peres SB, Henriques FS, Sertié RAL, Franco FO, Santos KB, Knobl P, Andreotti S, Shida CS, Neves RX, Farmer SR, Seelaender M, Lima FB, Batista Jr ML (2015) Pioglitazone treatment increases survival and prevents body weight loss in tumor-bearing animals: possible anti-cachectic effect. PLoS One 10:1–16. https://doi.org/10.1371/journal.pone.0122660
Bruheim S, Bruland OS, Breistol K, Maelandsmo GM, Fodstad Ø (2004) Human osteosarcoma xenografts and their sensitivity to chemotherapy. Pathol Oncol Res 10:133–141. https://doi.org/10.1007/bf03033741
Carrle D, Bielack SS (2006) Current strategies of chemotherapy in osteosarcoma. Int Orthop 30:445–451. https://doi.org/10.1007/s00264-006-0192-x
Chin L-H, Hsu S-P, Zhong W-B, Liang Y-C (2015) Combined treatment with troglitazone and lovastatin inhibited epidermal growth factor-induced migration through the downregulation of cysteine-rich protein 61 in human anaplastic thyroid cancer cells. PLoS One 10:1–16. https://doi.org/10.1371/journal.pone.0118674
Ciaramella V, Sasso FC, Di Liello R et al (2019) Activity and molecular targets of pioglitazone via blockade of proliferation, invasiveness and bioenergetics in human NSCLC. J Exp Clin Cancer Res 38:178–191
Eckland DA, Danhof M (2000) Clinical pharmacokinetics of pioglitazone. Exp Clin Endocrinol Diabetes 108:234–242
El-Sisi AE, Sokar SS, Salem TA, Abu Risha SE (2015) PPARgamma-dependent anti-tumor and immunomodulatory actions of pioglitazone. J Immunotoxicol 12:308–316. https://doi.org/10.3109/1547691X.2014.978055
Ghadiany M, Tabarraee M, Salari S, Haghighi S, Rezvani H, Ghasemi SN, Karimi-Sari H (2019) Adding oral Pioglitazone to standard induction chemotherapy of acute myeloid leukemia: a randomized clinical trial. Clin Lymphoma Myeloma Leuk 19:206–212. https://doi.org/10.1016/j.clml.2019.01.006
He H, Ni J, Huang JUN (2014) Molecular mechanisms of chemoresistance in osteosarcoma (Review). Oncol Lett 7:1352–1362. https://doi.org/10.3892/ol.2014.1935
Higuchi T, Sugisawa N, Miyake K et al (2019) Pioglitazone , an agonist of PPAR γ , reverses doxorubicin-resistance in an osteosarcoma patient-derived orthotopic xenograft model by downregulating P-glycoprotein expression. Biomed Pharmacother 118:4–8
Isakoff MS, Bielack SS, Meltzer P, Gorlick R (2015) Osteosarcoma: current treatment and a collaborative pathway to success. J Clin Oncol 33:3029–3035. https://doi.org/10.1200/JCO.2014.59.4895
Jaakkola T (2007) Pharmacokinetic interactions of pioglitazone. Helsinki
Jaffe N, Puri A, Gelderblom H (2013) Osteosarcoma: evolution of treatment paradigms. Sarcoma 2013:1–7. https://doi.org/10.1155/2013/203531
Kasper B, Ho AD, Egerer G (2005) A new therapeutic approach in patients with advanced sarcoma. Int J Clin Oncol 10:438–440. https://doi.org/10.1007/s10147-005-0514-9
Kawakami K, Kawakami M, Puri RK (2002) IL-13 receptor-targeted cytotoxin cancer therapy leads to complete eradication of tumors with the aid of phagocytic cells in nude mice model of human cancer. J Immunol 169:7119–7126. https://doi.org/10.4049/jimmunol.169.12.7119
Kostapanos MS, Elisaf MS, Mikhailidis DP (2013) Pioglitazone and cancer: angel or demon? Curr Pharm Des 19:4913–4929. https://doi.org/10.2174/13816128113199990294
Lu H, Waxman DJ (2005) Antitumor activity of methoxymorpholinyl doxorubicin: potentiation by cytochrome P450 3A metabolism. Mol Pharmacol 67:212–219
Lv S, Wang W, Wang H, Zhu Y, Lei C (2019) PPARγ activation serves as therapeutic strategy against bladder cancer via inhibiting PI3K-Akt signaling pathway. BMC Cancer 19:204–217
Makwana V, Dukie AS-A, Rudrawar S (2020) Investigating the impact of OGT inhibition on doxorubicin- and docetaxel-induced cytotoxicity in PC-3 and WPMY-1 cells. Int J Toxicol. https://doi.org/10.1177/1091581820948433
Martin JW, Squire JA, Zielenska M (2012) The genetics of osteosarcoma. Sarcoma 2012:1–11. https://doi.org/10.1155/2012/627254
Morrow JJ, Khanna C (2015) Osteosarcoma genetics and epigenetics: emerging biology and candidate therapies. Crit Rev Oncog 20:173–197
Papanagnou P, Stivarou T, Tsironi M (2016) Unexploited antineoplastic effects of commercially available anti-diabetic drugs. Pharm 9:24–44. https://doi.org/10.3390/ph9020024
Picci P, Serra M (2017) Doxorubicin-resistant osteosarcoma: novel therapeutic approaches in sight? Future Oncol 13:673–677
Posthumadeboer J, Van Royen BJ, Helder MN (2013) Mechanisms of therapy resistance in osteosarcoma: a review. Oncol Discov 1:1–15. https://doi.org/10.7243/2052-6199-1-8
Prost S, Relouzat F, Spentchian M, Ouzegdouh Y, Saliba J, Massonnet G, Beressi JP, Verhoeyen E, Raggueneau V, Maneglier B, Castaigne S, Chomienne C, Chrétien S, Rousselot P, Leboulch P (2015) Erosion of the chronic myeloid leukaemia stem cell pool by PPARgamma agonists. Nature 525:380–383. https://doi.org/10.1038/nature15248
Pushpakom S, Iorio F, Eyers PA, Escott KJ, Hopper S, Wells A, Doig A, Guilliams T, Latimer J, McNamee C, Norris A, Sanseau P, Cavalla D, Pirmohamed M (2019) Drug repurposing: progress, challenges and recommendations. Nat Rev Drug Discov 18:41–58. https://doi.org/10.1038/nrd.2018.168
Rajkumar T, Parija T YM (2010) Troglitazone modulates doxorubicin drug resistance through MDR1, p53, IL8 and ECOP. In: Kobayashi Foundation Award for Cancer Research in the 9th ACOS meeting in Gifu, Japan
Rajkumar T, Yamuna M (2008) Multiple pathways are involved in drug resistance to doxorubicin in an osteosarcoma cell line. Anti-Cancer Drugs 19:257–265
Ramachandran B, Jayavelu S, Murhekar K (2016) Repeated dose studies with pure Epigallocatechin-3-gallate demonstrated dose and route dependant hepatotoxicity with associated dyslipidemia. Toxicol Rep 3:336–345. https://doi.org/10.1016/j.toxrep.2016.03.001
Ramteke P, Deb A, Shepal V, Bhat MK (2019) Hyperglycemia associated metabolic and molecular alterations in cancer risk, progression, treatment, and mortality. Cancers (Basel) 11(9):1402. https://doi.org/10.3390/cancers11091402
Rickel K, Fang F, Tao J (2017) Molecular genetics of osteosarcoma. Bone 102:69–79. https://doi.org/10.1016/j.bone.2016.10.017
Rousselot P, Prost S, Guilhot J, Roy L, Etienne G, Legros L, Charbonnier A, Coiteux V, Cony-Makhoul P, Huguet F, Cayssials E, Cayuela JM, Relouzat F, Delord M, Bruzzoni-Giovanelli H, Morisset L, Mahon FX, Guilhot F, Leboulch P, on behalf of the French CML Group (2017) Pioglitazone together with imatinib in chronic myeloid leukemia: a proof of concept study. Cancer 123:1791–1799. https://doi.org/10.1002/cncr.30490
Saboo B, Kruljac I, Rahelić D (2015) Pioglitazone improves survival in patients with cancer: the hypothesis. Endocr Oncol Metab 1:24–33. https://doi.org/10.21040/eom/000004
Saiki M, Hatta Y, Yamazaki T, Itoh T, Enomoto Y, Takeuchi J, Sawada U, Aizawa S, Horie T (2006) Pioglitazone inhibits the growth of human leukemia cell lines and primary leukemia cells while sparing normal hematopoietic stem cells. Int J Oncol 29:437–443
Shen Z, Reed JR, Creighton M, Liu DQ, Tang YS, Hora DF, Feeney W, Szewczyk J, Bakhtiar R, Franklin RB, Vincent SH (2003) Identification of novel metabolites of pioglitazone in rat and dog. Xenobiotica 33:499–509. https://doi.org/10.1080/0049825031000085951
Venkatakrishnan K, Von Moltke LL, Greenblatt DJ (2001) Human drug metabolism and the cytochromes P450: application and relevance of in vitro models. J Clin Pharmacol 41:1149–1179. https://doi.org/10.1177/00912700122012724
Wagner ER, He B, Chen L et al (2010) Therapeutic implications of PPAR γ in human osteosarcoma. PPAR Res 2010:1–16. https://doi.org/10.1155/2010/956427
Wang C, Wang J, Bai P (2011) Troglitazone induces apoptosis in gastric cancer cells through the NAG-1 pathway. Mol Med Rep 4:93–97. https://doi.org/10.3892/mmr.2010.381
Winkler K, Bielack SS, Delling G, et al (1993) Treatment of osteosarcoma: experience of the Cooperative Osteosarcoma Study Group (COSS). 62:269–77
Acknowledgments
The authors wish to thank M. Vijayavel for his technical assistance.
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Aparna Natarajan stipend [JRF and SRF] was supported by the Department of Biotechnology (DBT), Government of India (BT/PR1457/2011).
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All the in vitro and in vivo data were generated in-house and no paper mill was used. TR conceptualized, supervised the study, and analyzed the data. AN carried out all the in vitro, in vivo, and HPLC experiments, analyzed the data, and drafted the manuscript. BR performed and supervised the animal studies at the departmental animal house facility. GG supervised the HPLC study. SJ assisted with HPLC experiments at the departmental proteomics facility. SS evaluated the H and E slides. All authors discussed the results and contributed and approved the final manuscript.
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All animal studies were carried out with prior approval from the Institutional Animal Ethics Committee (IAEC) of Cancer Institute (W.I.A), India. Care of animals complied according to CPCSEA (Committee for the purpose of Control and Supervision of Experiments on Animals) guidelines, Government of India.
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Natarajan, A., Ramachandran, B., Gopisetty, G. et al. Pioglitazone modulates doxorubicin resistance in a in vivo model of drug resistant osteosarcoma xenograft. Naunyn-Schmiedeberg's Arch Pharmacol 394, 361–371 (2021). https://doi.org/10.1007/s00210-020-01982-3
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DOI: https://doi.org/10.1007/s00210-020-01982-3