The role of solute carrier (SLC) transporters in actinomycin D pharmacokinetics in paediatric cancer patients
Actinomycin D is used for treatment of paediatric cancers; however, a large inter-patient pharmacokinetic (PK) variability and hepatotoxicity are significant limitations to its use and warrant further investigation. Elimination of actinomycin D may be mediated by transporters, as the drug does not seem to undergo significant metabolism. We investigated the role of solute carrier (SLC) transporters in actinomycin D PK.
Fourteen key SLCs were screened through probe substrate uptake inhibition by actinomycin D in HEK293 cells. Uptake of actinomycin D was further studied in candidate SLCs by measuring intracellular actinomycin D using a validated LCMS assay. Pharmacogenetic analysis was conducted for 60 patients (Clinical trial: NCT00900354), who were genotyped for SNPs for OAT4 and PEPT2.
OAT4, OCT2, OCT3 and PEPT2 showed significantly lower probe substrate uptake (mean ± SD 75.0 ± 3.5% (p < 0.0001), 74.8 ± 11.2% (p = 0.001), 81.2 ± 14.0% (p = 0.0083) and 70.7 ± 5.7% (p = 0.0188)) compared to that of control. Intracellular accumulation of actinomycin D was greater compared to vector control in OAT4-transfected cells by 1.5- and 1.4-fold at 10 min (p = 0.01) and 20 min (p = 0.03), and in PEPT2-transfected cells by 1.5- and 1.7-fold at 10 min (p = 0.047) and 20 min (p = 0.043), respectively. Subsequent clinical study did not find a significant association between OAT4 rs11231809 and PEPT2 rs2257212 genotypes, and actinomycin D PK parameters such as clearance (CL) and volume of distribution (Vd).
Transport of actinomycin D was mediated by OAT4 and PEPT2 in vitro. There was a lack of clinical significance of OAT4 and PEPT2 genotypes as predictors of actinomycin D disposition in paediatric cancer patients.
KeywordsActinomycin D SLC transporters Pharmacokinetics Pharmacogenetics Cancer
This work was supported by the University of Sydney, Australia, Cancer Research UK and the Experimental Cancer Medicine Centre Network.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Fernandez CV, Mullen EA, Chi YY, Ehrlich PF, Perlman EJ, Kalapurakal JA, Khanna G, Paulino AC, Hamilton TE, Gow KW, Tochner Z, Hoffer FA, Withycombe JS, Shamberger RC, Kim Y, Geller JI, Anderson JR, Grundy PE, Dome JS (2018) Outcome and prognostic factors in stage III favorable-histology Wilms tumor: a report from the Children’s Oncology Group Study AREN0532. J Clin Oncol 36(3):254–261CrossRefGoogle Scholar
- 3.Gaspar N, Hawkins DS, Dirksen U, Lewis IJ, Ferrari S, le Deley MC, Kovar H, Grimer R, Whelan J, Claude L, Delattre O, Paulussen M, Picci P, Sundby Hall K, van den Berg H, Ladenstein R, Michon J, Hjorth L, Judson I, Luksch R, Bernstein ML, Marec-Bérard P, Brennan B, Craft AW, Womer RB, Juergens H, Oberlin O (2015) Ewing sarcoma: current management and future approaches through collaboration. J Clin Oncol 33(27):3036–3046CrossRefGoogle Scholar
- 10.Veal GJ, Cole M, Errington J, Parry A, Hale J, Pearson AD, Howe K, Chisholm JC, Beane C, Brennan B, Waters F, Glaser A, Hemsworth S, McDowell H, Wright Y, Pritchard-Jones K, Pinkerton R, Jenner G, Nicholson J, Elsworth AM, Boddy AV, Kingdom Children's Cancer Study Group Pharmacology Working Group (2005) Pharmacokinetics of dactinomycin in a pediatric patient population: a United Kingdom Children’s Cancer Study Group Study. Clin Cancer Res 11(16):5893–5899CrossRefGoogle Scholar
- 12.Skolnik JM, Herman JL, Adamson PC (2008) Identification of metabolites of actinomycin-D in vivo and in vitro. in 37th Annual Meeting, American College of Clinical Pharmacology. J Clin Pharmacol Philadelphia, Pennsylvania 48:1128Google Scholar
- 22.Zhou F, Zhu L, Wang K, Murray M (2017) Recent advance in the pharmacogenomics of human Solute Carrier Transporters. Adv Drug Deliv Rev 116:21–36Google Scholar
- 23.UniProt (2017) the universal protein knowledgebase. Nucleic Acids Res 45(D1):D158–d169Google Scholar
- 27.Lu X, Chan T, Zhu L, Bao X, Velkov T, Zhou QT, Li J, Chan HK, Zhou F (2018) The inhibitory effects of eighteen front-line antibiotics on the substrate uptake mediated by human organic anion/cation transporters, organic anion transporting polypeptides and oligopeptide transporters in in vitro models. Eur J Pharm Sci 115:132–143CrossRefGoogle Scholar
- 30.Poirier A, Belli S, Funk C, Otteneder MB, Portmann R, Heinig K, Prinssen E, Lazic SE, Rayner CR, Hoffmann G, Singer T, Smith DE, Schuler F (2012) Role of the intestinal peptide transporter PEPT1 in oseltamivir absorption: in vitro and in vivo studies. Drug Metab Dispos 40(8):1556–1565CrossRefGoogle Scholar
- 40.Ortega JA, Donaldson SS, Ivy SP, Pappo A, Maurer HM (1997) Venoocclusive disease of the liver after chemotherapy with vincristine, actinomycin D, and cyclophosphamide for the treatment of rhabdomyosarcoma. A report of the Intergroup Rhabdomyosarcoma Study Group. Childrens Cancer Group, the Pediatric Oncology Group, and the Pediatric Intergroup Statistical Center. Cancer 79(12):2435–2439CrossRefGoogle Scholar
- 41.Green DM, Breslow NE, Beckwith JB, Finklestein JZ, Grundy PE, Thomas PR, Kim T, Shochat SJ, Haase GM, Ritchey ML, Kelalis PP, D'Angio GJ (1998) Comparison between single-dose and divided-dose administration of dactinomycin and doxorubicin for patients with Wilms’ tumor: a report from the National Wilms’ Tumor Study Group. J Clin Oncol 16(1):237–245CrossRefGoogle Scholar
- 42.Green DM, Breslow NE, Beckwith JB, Finklestein JZ, Grundy P, Thomas PR, Kim T, Shochat S, Haase G, Ritchey M, Kelalis P, D'Angio GJ (1998) Effect of duration of treatment on treatment outcome and cost of treatment for Wilms’ tumor: a report from the National Wilms’ Tumor Study Group. J Clin Oncol 16(12):3744–3751CrossRefGoogle Scholar
- 44.Tournade MF, Com-Nougué C, de Kraker J, Ludwig R, Rey A, Burgers JM, Sandstedt B, Godzinski J, Carli M, Potter R, Zucker JM, International Society of Pediatric Oncology Nephroblastoma Trial and Study Committee (2001) Optimal duration of preoperative therapy in unilateral and nonmetastatic Wilms’ tumor in children older than 6 months: results of the Ninth International Society of Pediatric Oncology Wilms’ Tumor Trial and Study. J Clin Oncol 19(2):488–500CrossRefGoogle Scholar
- 45.Arndt CA et al (2009) Vincristine, actinomycin, and cyclophosphamide compared with vincristine, actinomycin, and cyclophosphamide alternating with vincristine, topotecan, and cyclophosphamide for intermediate-risk rhabdomyosarcoma: children’s oncology group study D9803. J Clin Oncol 27(31):5182–5188CrossRefGoogle Scholar
- 46.Graf N, van Tinteren H, Bergeron C, Pein F, van den Heuvel-Eibrink MM, Sandstedt B, Schenk JP, Godzinski J, Oldenburger F, Furtwängler R, de Kraker J (2012) Characteristics and outcome of stage II and III non-anaplastic Wilms’ tumour treated according to the SIOP trial and study 93-01. Eur J Cancer 48(17):3240–3248CrossRefGoogle Scholar
- 47.Pritchard-Jones K, Bergeron C, de Camargo B, van den Heuvel-Eibrink MM, Acha T, Godzinski J, Oldenburger F, Boccon-Gibod L, Leuschner I, Vujanic G, Sandstedt B, de Kraker J, van Tinteren H, Graf N (2015) Omission of doxorubicin from the treatment of stage II-III, intermediate-risk Wilms’ tumour (SIOP WT 2001): an open-label, non-inferiority, randomised controlled trial. Lancet 386(9999):1156–1164CrossRefGoogle Scholar
- 48.Skolnik JM, Herman JL, Adamson PC (2008) Identification of metabolites of actinomycin-D in vivo and in vitro. J Clin Pharmacol 48(9):1128Google Scholar
- 64.Lee YS, Kim BH, Kim BC, Shin A, Kim JS, Hong SH, Hwang JA, Lee JA, Nam S, Lee SH, Bhak J, Park JW (2015) SLC15A2 genomic variation is associated with the extraordinary response of sorafenib treatment: whole-genome analysis in patients with hepatocellular carcinoma. Oncotarget 6(18):16449–16460PubMedPubMedCentralGoogle Scholar
- 66.Yao S, Sucheston LE, Zhao H, Barlow WE, Zirpoli G, Liu S, Moore HCF, Thomas Budd G, Hershman DL, Davis W, Ciupak GL, Stewart JA, Isaacs C, Hobday TJ, Salim M, Hortobagyi GN, Gralow JR, Livingston RB, Albain KS, Hayes DF, Ambrosone CB (2014) Germline genetic variants in ABCB1, ABCC1 and ALDH1A1, and risk of hematological and gastrointestinal toxicities in a SWOG phase III trial S0221 for breast cancer. Pharmacogenomics J 14(3):241–247CrossRefGoogle Scholar
- 73.Moscow JA, Gong M, He R, Sgagias MK, Dixon KH, Anzick SL, Meltzer PS, Cowan KH (1995) Isolation of a gene encoding a human reduced folate carrier (RFC1) and analysis of its expression in transport-deficient, methotrexate-resistant human breast cancer cells. Cancer Res 55(17):3790–3794PubMedGoogle Scholar
- 74.Lopez-Lopez E, Ballesteros J, Piñan MA, Sanchez de Toledo J, Garcia de Andoin N, Garcia-Miguel P, Navajas A, Garcia-Orad A (2013) Polymorphisms in the methotrexate transport pathway: a new tool for MTX plasma level prediction in pediatric acute lymphoblastic leukemia. Pharmacogenet Genomics 23(2):53–61CrossRefGoogle Scholar