Cancer Chemotherapy and Pharmacology

, Volume 57, Issue 1, pp 7–14

Aplidin reduces growth of anaplastic thyroid cancer xenografts and the expression of several angiogenic genes

  • Ann M. Straight
  • Kevin Oakley
  • Russell Moores
  • Andrew J. Bauer
  • Aneeta Patel
  • R. Michael Tuttle
  • J. Jimeno
  • Gary L. Francis
Original Article


Background Anaplastic thyroid cancer (ATC) is one of the most aggressive and highly lethal human cancers. Median survival after diagnosis is 4–6 months despite available radiotherapy and chemotherapy. Additional treatments are needed for ATC. Vascular endothelial growth factor (VEGF) is a potent angiogenic stimulus, which is expressed by ATC. Previously, anti-VEGF antibody was used to block VEGF-dependent angiogenesis in ATC xenografts. This treatment induced partial (56%) but not complete tumor regression. Aplidin (APLD) is a marine derived antitumor agent currently in phase II clinical studies. Multiple activities of this compound have been described which likely contribute to its antiproliferative effect. Notably, APLD has been shown to have antiangiogenic properties which include: inhibition of VEGF secretion, reduction in the synthesis of the VEGF receptor (FLT-1), and blockade of matrix metalloproteinase production by endothelial cells. We hypothesized that Aplidin, with its broad spectrum of action and antiangiogenic properties, would be a potentially effective drug against ATC. Methods Thirty BALB/c nu/nu mice were injected with ATC cells (ARO-81, 1×106) and allowed to implant for 3 weeks. Animals were randomized to receive daily intraperitoneal injections of vehicle, low dose (0.5 mg/kg/day), or high dose (1.0 mg/kg/day) APLD. After 3 days, the animals were killed and the tumors were removed, weighed, and divided for RNA and protein analyses. Results APLD significantly reduced ATC xenograft growth (low dose, 20% reduction, P=0.01; high dose, 40% reduction, P<0.001). This was associated with increased levels of apoptosis related proteins polyadenosylribose polymerase 85 (PARP-85, 75% increase, P=0.024) and caspase 8 (greater than fivefold increase, P=0.03). APLD treatment was further associated with lost or reduced expression of several genes that support angiogenesis to include: VEGF, hypoxia inducible factor 1(HIF-1), transforming growth factor-beta (TGFβ), TGFβ receptor 2 (TGFβR2), melanoma growth stimulating factor 1 (GRO1), cadherin, and vasostatin. Conclusions This data supports the hypothesis that APLD may be an effective adjunctive therapy against ATC. The demonstrated molecular impact against angiogenic related genes specifically supports future strategies combining APLD with VEGF interacting agents.


Thyroid cancer Aplidin Angiogenesis 


  1. 1.
    Aardal S, Helle KB (1992) The vasoinhibitory activity of bovine chromogranin A fragment (vasostatin) and its independence of extracellular calcium in isolated segments of human blood vessels. Regul Pept 41:9–18CrossRefPubMedGoogle Scholar
  2. 2.
    Ain KB, Egorin MJ, DeSimone PA (2000) Treatment of anaplastic thyroid carcinoma with paclitaxel: phase 2 trial using 96-h infusion Collaborative Anaplastic Thyroid Cancer Health Intervention Trials (CATCHIT) Group. Thyroid 10:587–594PubMedGoogle Scholar
  3. 3.
    Ain KB, Tofiq S, Taylor KD (1996) Antineoplastic activity of taxol against human anaplastic thyroid carcinoma cell lines in vitro and in vivo. J Clin Endocrinol Metab 81:3650–3653CrossRefPubMedGoogle Scholar
  4. 4.
    Bauer AJ, Patel A, Terrell R, Doniparthi K, Saji M, Ringel M, Tuttle RM, Francis G (2003) Vascular endothelial growth factor monoclonal antibley (VEGF-MAb) inhibits growth of papillary thyroid cancer xenografts. Ann Clin Lab Sci 33:192–199PubMedGoogle Scholar
  5. 5.
    Bauer AJ, Terrell R, Doniparthi NK, Patel A, Tuttle RM, Saji M, Ringel M, Francis G (2002) Vascular endothelial growth factor monoclonal antibody inhibits growth of anaplastic thyroid cancer xenografts in nude mice. Thyroid 12:953–961CrossRefPubMedGoogle Scholar
  6. 6.
    Braga-Basaria M, Ringel M (2003) Beyond radioiodine: a review of potential new therapeutic approaches for thyroid cancer. J Clin Endocrinol Metab 88(5):1947–1960CrossRefPubMedGoogle Scholar
  7. 7.
    Bresters D, Broekhuizen AJF, Kaaijk P, Faircloth GT, Jimeno J, Kaspers GJL (2003) In vitro cytotoxicity of aplidin and crossresistance with other cytotoxic drugs in childhood leukemic and normal bone marrow and blood samples: a rational basis for clinical development. Leukemia 17:1338–1343CrossRefPubMedGoogle Scholar
  8. 8.
    Broggini M, Marchini SV, Galliera E, Borsotti P, Taraboletti G, Erba E, Sironi M, Jimeno J, Faircloth GT, Giavazzi R, D’Incalci M (2003) Aplidine, a new anticancer agent of marine origin, inhibits vascular endothelial growth factor (VEGF) secretion and blocks VEGF-VEGFR-1 (flt- 1) autocrine loop in human leukemia cells MOLT-4. Leukemia 17:52–59CrossRefPubMedGoogle Scholar
  9. 9.
    Cano A, Perez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG, Portillo F, Nieto MA (2000) The transcription factor Snail controls epithelial-mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2:76–83CrossRefPubMedGoogle Scholar
  10. 10.
    Cardenas F, Thornmann M, Feliz M, Caba J, Lloyd-Williams P, Giralt E (2001) Conformational analysis of dehydrodidemnin B (Aplidine) by NMR spectroscopy and molecular mechanics/dynamics calculations. J Org Chem 66:4580–4584CrossRefPubMedGoogle Scholar
  11. 11.
    Carmeliet P, Jain R (2000) Angiogenesis in cancer and other diseases. Nature 407:249–256CrossRefPubMedGoogle Scholar
  12. 12.
    Celli N, Gallardo AM, Rossi C, Zucchetti M, D’Incalci M, Rotilio D (1999) Analysis of aplidine (dehydrodidemnin B), a new marine-derived depsipeptide, in rat biological fluids by liquid chromatography-tandem mass spectrometry. J Chromatogr B Biomed Sci Appl 731:335–343CrossRefPubMedGoogle Scholar
  13. 13.
    Crampton SL, Adams EG, Kuentzel SL, Li LH, Badiner G, Bhuyan BK (1984) Biochemical and cellular effects of didemnins A and B. Cancer Res 44:1796–1801PubMedGoogle Scholar
  14. 14.
    Crew CM, Collins JL, Lane WS, Snapper ML, Schreiber SL (1994) GTP-dependent binding of the antiproliferative agen didenin to elongation factor 1 alpha. J Biol Chem 269:15411–15415PubMedGoogle Scholar
  15. 15.
    Depenbrock H, Peter R, Faircloth GT, Manzanares I, Jimeno J, Hanauske AR (1998) In vitro activity of aplidine, a new marine-derived anti-cancer compound, on freshly explanted clonogenic human tumour cells and haematopoietic precursor cells. Br J Cancer 78:739–744PubMedGoogle Scholar
  16. 16.
    Dvorak HF, Nagy JA, Feng D, Brown LF, Dvorak AM (1999) Vascular permeability factor/ vascularendothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Microbiol Immunol 237:97–132PubMedGoogle Scholar
  17. 17.
    Dziba JM, Marcinek R, Venkataraman G, Robinson JA, Ain KB (2002) Combretastatin A4 phosphate has primary antineoplastic activity against human anaplastic thyroid carcinoma cell lines and xenograft tumors. Thyroid 12 (12):1063–1070CrossRefPubMedGoogle Scholar
  18. 18.
    Erba E, Bassano L, Di Liberti G, Muradore I, Chiorino G, Ubezio P, Vignati S, Codegoni A, Desiderio MA, Faircloth G, Jimeno J, D’Incalci M (2002) Cell cycle phase perturbations and apoptosis in tumour cells induced by aplidine. Br J Cancer 86:1510–1517CrossRefPubMedGoogle Scholar
  19. 19.
    Fenton C, Patel A, Dinauer C, Robie DK, Tuttle RM, Francis GL (2000) The expression of vascular endothelial growth factor and the type 1 vascular endothelial growth factor receptor correlate with the size of papillary thyroid carcinoma in children and young adults. Thyroid 10: 349–357PubMedCrossRefGoogle Scholar
  20. 20.
    Gasparri A (1997) Chromogranin A fragments modulate cell adhesion. Identification and characterization of a pro-adhesive domain. J Biol Chem 272:20835–20843CrossRefPubMedGoogle Scholar
  21. 21.
    Geldof AA, Mastbergen SC, Henrar RE, Faircloth GT (1999) Cytotoxicity and neurocytotoxicity of new marine anticancer agents evaluated using in vitro assays. Cancer Chemother Pharmacol 44:312–318CrossRefPubMedGoogle Scholar
  22. 22.
    Gerber H, McMurtrey A, Kowalski J, Yan M, Keyt BA, Dixit V, Ferrara N (1998) Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3’-kinase/Akt signal transduction pathway. Requirement for the Flk-1/KDR activation. J Biol Chem 273:30336–30343CrossRefPubMedGoogle Scholar
  23. 23.
    Hama Y, Shimizu T, Hosaka S, Sugenoya A, Usuda N (1997) Therapeutic efficacy of the angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470; AGM-1470) for human anaplastic thyroid carcinoma in nude mice. Exp Toxicol Pathol 49:239–247PubMedGoogle Scholar
  24. 24.
    Hammani K, Blakis A, Morsette D, et al. (1996) Structure and characterization of the human tissue inhibitor of metalloproteinases-2 gene. J Biol Chem 271:25498–25505CrossRefPubMedGoogle Scholar
  25. 25.
    Huang SM, Lee JC, Wu TJ, Chow NH (2001) Clinical relevance of vascular endothelial growth factor for thyroid neoplasms. World J Surg 25:302–306CrossRefPubMedGoogle Scholar
  26. 26.
    Imamura Y, Jin L, Grande JP, Li CY, Zheng TR, Erickson LA, Lloyd R (1998) Analysis of TGF-B and TGF-B-RII in thyroid neoplasms from the United States, Japan, and China. Endocr Pathol 9(3):209–216PubMedCrossRefGoogle Scholar
  27. 27.
    Ishiwata I, Sudo T, Kiguchi K, Ishikawa H (1999) Tumor angiogenesis factors produced by cancer cells. Hum Cell 12:37–46PubMedGoogle Scholar
  28. 28.
    Jimeno J, Lopez-Martin J, Ruiz-Casado A, Izquierdo M, Scheuer P, Rinehart K (2004) Progress in the clinical development of new marine-derived anticancer compounds. Anticancer Drugs 15:321–329CrossRefPubMedGoogle Scholar
  29. 29.
    Lennard CM, Patel A, Wilson J, Reinhardt B, Tuman C, Fenton C, Blair E, Francis GL, Tuttle RM (2001) Intensity of vascular endothelial growth factor expression is associated with increased risk of recurrence and decreased disease-free survival in papillary thyroid cancer. Surgery 129:552–558CrossRefPubMedGoogle Scholar
  30. 30.
    Lessin LS, Min M (2000) Chemotherapy of anaplastic thyroid cancer. In: Wartofsky L (ed) Thyroid cancer: a comprehensive guide to clinical management. Humana, Totowa, pp 337–340Google Scholar
  31. 31.
    Lo CY, Lam KY, Wan KY (1999) Anaplastic carcinoma of the thyroid. Am J Surg 177:337–339CrossRefPubMedGoogle Scholar
  32. 32.
    Lugardon K (2000) Antibacterial and antifungal activities of vasostatin-1, the N-terminal fragment of chromogranin A. J Biol Chem 275:10745–10753CrossRefPubMedGoogle Scholar
  33. 33.
    Marchini S, Chiorino G, Faircloth GT, D’Incalci M (2002) Changes in gene expression profile induced by the anticancer agent Aplidine in Molt-4 leukemic cell lines. J Biol Regul Homeost Agents 16:241–248PubMedGoogle Scholar
  34. 34.
    Mastbergen SC, Duivenvoorden I, Versteegh RT, Geldof AA (2000) Cell cycle arrest and clonogenic tumor cell kill by divergent chemotherapeutic drugs. Anticancer Res 20:1833–1838PubMedGoogle Scholar
  35. 35.
    National Cancer Institute, SEER Program (1999) Cancer Incidence and Survival among Children and Adolescents: United States SEER Program 1975–1995. NIH, BethesdaGoogle Scholar
  36. 36.
    Noda K, Ishida S, Inoue M, Obata K, Oguchi Y, Okada Y, Ikeda E (2003) Production and activation of matrix metalloproteinase-2 in proliferative diabetic retinopathy. Invest Ophthal Vis Sci 44:2163–2170CrossRefPubMedGoogle Scholar
  37. 37.
    Nuijen B, Bouma M, Henrar RE, Floriano P, Jimeno JM, Talsma H, Kettenes-van den Bosch JJ, Heck AJ, Bult A, Beijnen JH (2000) Pharmaceutical development of a parenteral lyophilized formulation of the novel antitumor agent aplidine. PDA J Pharm Sci Technol 54:193–208PubMedGoogle Scholar
  38. 38.
    Nuijen B, Bouma M, Henrar RE, Manada C, Bult A, Beijnen JH (1999) Compatibility and stability of aplidine, a novel marine-derived depsipeptide antitumor agent, in infusion devices, and its hemolytic and precipitation potential upon iv administration. Anticancer Drugs 10:879–887PubMedCrossRefGoogle Scholar
  39. 39.
    Raymond E, Ady-Vago N, Baudin E, Ribrag V, Faivre S, Lecot F (2000) A phase 1 and pharmacokinetic study of aplidin given as a 24-h continuous infusion every other week in patients with solid tumors and lymphoma. Clin Cancer Res 6(Suppl):4510–4515Google Scholar
  40. 40.
    Rinehart KL (2000) Antitumor compounds from tunicates. Med Res Rev 20:1–27CrossRefPubMedGoogle Scholar
  41. 41.
    Sager R (1990) GRO as a cytokine. In: Oppenheim J (ed) Molecular and cellular biology of cytokines. Wiley-Liss, New York, pp 327–332Google Scholar
  42. 42.
    Sparidans RW, Kettenes-van den Bosch JJ, van Tellingen O, Nuyen B, Henrar RE, Jimeno JM, Faircloth G, Floriano P, Rinehart KL, Beijnen JH (1999) Bioanalysis of aplidine, a new marine antitumoral depsipeptide, in plasma by high-performance liquid chromatography after derivatization with trans-hydrazino-2-stilbazole. J Chromatogr B Biomed Sci Appl 729:43–53CrossRefPubMedGoogle Scholar
  43. 43.
    Straight A, Patel A, Jimeno J, Tuttle RM, Francis GL (2003) Aplidine Reduces In Vivo and In Vitro Growth of Anaplastic Thytoid Cancer. The Endocrine Society, PhiladelphiaGoogle Scholar
  44. 44.
    Taraboletti G, Poli M, Manenti L, Borsotti P, Broggini M, D’ Incalci M, Ribatti D, Giavazzi R (2004) Antiangiogenic activity of aplidine, a new agent of marine origin. Br J Cancer 90:2418–2424PubMedGoogle Scholar
  45. 45.
    Tennvall J, Lundell G, Hallquist A, Wahlberg P, Wallin G, Tibblin S (1994) Combined doxorubicin, hyperfractionated radiotherapy, and surgery in anaplastic thyroid carcinoma Report on two protocols The Swedish Anaplastic Thyroid Cancer Group. Cancer 74:1348–1354PubMedCrossRefGoogle Scholar
  46. 46.
    Tuttle RM, Fleisher M, Francis GL, Robbins RJ (2002) Serum vascular endothelial growth factor levels are elevated in metastatic differentiated thyroid cancer but not increased by short-term TSH stimulation. J Clin Endocrinol Metab 87:1737–1742CrossRefPubMedGoogle Scholar
  47. 47.
    Tuttle RM, Patel A, Francis G, Davis S, Kopecky KJ, Lushnikov E, Abrosimov A, Troshin V, Tsyb A, Fenton C (2000) Vascular endothelial growth factor (VEGF) and Type 1 VEGF receptor (Flt-1) are highly expressed in Russian papillary thyroid carcinomas. In: 12th International Thyroid Congress, Kyoto, JapanGoogle Scholar
  48. 48.
    Urdiales JL, Morata P, De Castro IN, Sanchez-Jimenez F (1996) Antiproliferative effect of dehydrodidemnin B (DDB), a depsipeptide isolated from the Mediterranean tunicates. Cancer Lett 102:31–37CrossRefPubMedGoogle Scholar
  49. 49.
    West J, Munoz-Antonia T, Johnson JG, Klotch D, Muro-Cacho CA (2000) Transforming growth factor-beta type II receptors and Smad proteins in follicular thyroid tumors. Laryngoscope 110(8):1323–1327CrossRefPubMedGoogle Scholar
  50. 50.
    Zachary I (1998) Vascular endothelial growth factor. Int J Biochem Cell Biol 30:1169–1174CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Ann M. Straight
    • 1
    • 2
  • Kevin Oakley
    • 2
  • Russell Moores
    • 2
  • Andrew J. Bauer
    • 1
    • 2
  • Aneeta Patel
    • 2
  • R. Michael Tuttle
    • 3
  • J. Jimeno
    • 4
  • Gary L. Francis
    • 1
    • 2
  1. 1.Department of PediatricsWalter Reed Army Medical CenterWashingtonUSA
  2. 2.Department of PediatricsUniformed Services UniversityBethesdaUSA
  3. 3.Section of EndocrinologyMemorial Sloan Kettering Cancer CenterNew YorkUSA
  4. 4.PharmaMar SAU R&D Colmenar ViejoMadridSpain

Personalised recommendations