Evaluation of DNA damage in a Her2+ cell line induced by an Auger-emitting immunoconjugate

Document Type : Original Article

Authors

Radiation Application Research School, Nuclear Science and Technology Research Institute, Tehran, Iran

Abstract

Introduction: Auger electron based radioimmunotherapy (RIT) using 111In-DOTA-trastuzumab (111In-DOTA-antiHer2) feasibility was studied in vitro on a HER2/neu positive cell line, SkBr3.
Methods:111In-DOTA-antiHer2 was prepared according to the optimized conditions followed by quality control tests including radiochemical purity; immunoreactivity). SkBr3 as a HER2/neu positive cell line was used to determine the degree of internalization and cell viability of 111In-DOTA-antiHer2. For comet assay freshly cultivated SkBr3 cells incubated with 111In-DOTA-antiHer2 in 3.7 and 7.4 MBq doses at 37ºC for 24h.
Results:111In-DOTA-antiHer2 (>95% radiochemical purity; >79% immunoreactivity) demonstrated significant internalization in SKBr3 cells in 24 h. no significant cell viability difference observed for 111In-DOTA-antiHer2 and 111In cation treatments. Comet assay at 3.7 MBq demonstrated no significant DNA damage, while at 7.4 MBq dose DNA damage observed at least 16% more than control In-111 chloride after 24 h.
Conclusion: Although the internalization of 111In-DOTA-antiHer2 was approved in this study, however, lack of cell death and slight DNA breakage for 111In-DOTA-antiHer2 treatment suggests absence of nucleus entry which is essential for demonstration of DNA damage according to Auger electron range at cellular level.

Keywords

Main Subjects

References

  1. Welsh JS. Beta decay in science and medicine. Am J Clin Oncol. 2007 Aug;30(4):437-9.
  2. Seidl C. Radioimmunotherapy with α-particle-emitting radionuclides. Immunotherapy. 2014;6(4):431-58.
  3. Li L, Quang TS, Gracely EJ, Kim JH, Emrich JG, Yaeger TE, Jenrette JM, Cohen SC, Black P, Brady LW. A Phase II study of anti-epidermal growth factor receptor radioimmunotherapy in the treatment of glioblastoma multiforme. J Neurosurg. 2010 Aug;113(2):192-8.
  4. Quang TS, Brady LW. Radioimmunotherapy as a novel treatment regimen: 125I-labeled monoclonal antibody 425 in the treatment of high-grade brain gliomas. Int J Radiat Oncol Biol Phys. 2004 Mar 1;58(3):972-5.
  5. Kairemo KJ, Ramsay HA, Paavonen TK, Jääskelä-Saari HA, Tagesson M, Ljunggren K, Strand SE. Biokinetics of indium-111-bleomycin complex in head and neck cancer--implementations for radiochemotherapy. Cancer Detect Prev. 1997;21(1):83-90.
  6. Limouris GS, Lyra M, Skarlos D, Hatzioannou H, Gouliamos A, Moulopoulou A, Vlahos L. Auger and conversion electron therapy with In-111 pentetreotide in hepatocellural carcinoma. In: Bergmann H, Kohn H, Sinzinger H. Radioactive Isotopes in Clinical Medicine and Research XXIII. Basel/Switzerland: Brikhauser Verlag; 1999. p. 551-554.
  7. Steinsträsser A, Schwarz A, Kuhlmann L, Seidel L, Niemann E, Bosslet K. Preclinical and clinical testing of radiolabelled monoclonal antibodies for immunoscintigraphy. Dev Biol Stand. 1990;71:137-45.
  8. Sastry KSR, Rao DV. Dosimetry of low energy electrons. In: Rao DV, Chandra R, Graham MC, eds. Physics of Nuclear Medicine: Recent Advances. Woodbury, NY: American Institute of Physics; 1984. p. 169–208.
  9. Khorami-Moghadam A, Jalilian AR, Yavari K, Alirezapour B, Mazidi M, Mirzaii M. Synthesis, radiolabeling and quality control of 111In-DOTA-bevacizumab for radioimmunoscintigraphy of VEGF receptors. Radiochimica Acta. 2013;101:577-584.
  10. Alirezapour B, Abbas Abadi S, Kertesz I, Mohammadnejad J, Jalilian AR, Maadi E. [111In]-DOTA-anti-MUC1 for SPECT applications, the production and quality control. Eur J Nucl Med Mol Imaging. 2015;42(Suppl 1):S461.
  11. Leyton JV, Gao C, Williams B, Keating A, Minden M, Reilly RM. A radiolabeled antibody targeting CD123(+) leukemia stem cells - initial radioimmunotherapy studies in NOD/SCID mice engrafted with primary human AML. Leuk Res Rep. 2015 Sep 18;4(2):55-9.
  12. Razumienko EJ, Chen JC, Cai Z, Chan C, Reilly RM. Dual-Receptor-Targeted Radioimmunotherapy of Human Breast Cancer Xenografts in Athymic Mice Coexpressing HER2 and EGFR Using 177Lu- or 111In-Labeled Bispecific Radioimmunoconjugates. J Nucl Med. 2016 Mar;57(3):444-52.
  13. Wicki A, Wild D, Prêtre V, Mansi R, Orleth A, Reubi JC, Rochlitz C, Mamot C, Mäcke HR, Christofori G. Synergism of peptide receptor-targeted Auger electron radiation therapy with anti-angiogenic compounds in a mouse model of neuroendocrine tumors. EJNMMI Res. 2014 Feb 16;4(1):9.
  14. Cornelissen B, Darbar S, Hernandez R, Kersemans V, Tullis I, Barber PR, Smart S, Vojnovic B, Reilly R, Vallis KA. ErbB-2 blockade and prenyltransferase inhibition alter epidermal growth factor and epidermal growth factor receptor trafficking and enhance (111)In-DTPA-hEGF Auger electron radiation therapy. J Nucl Med. 2011 May;52(5):776-83.
  15. Costantini DL, McLarty K, Lee H, Done SJ, Vallis KA, Reilly RM. Antitumor effects and normal-tissue toxicity of 111In-nuclear localization sequence-trastuzumab in athymic mice bearing HER-positive human breast cancer xenografts. J Nucl Med. 2010 Jul;51(7):1084-91
  16. Kersemans V, Cornelissen B, Minden MD, Brandwein J, Reilly RM. Drug-resistant AML cells and primary AML specimens are killed by 111In-anti-CD33 monoclonal antibodies modified with nuclear localizing peptide sequences. J Nucl Med. 2008 Sep;49(9):1546-54.
  17. Bailey KE, Costantini DL, Cai Z, Scollard DA, Chen Z, Reilly RM, Vallis KA. Epidermal growth factor receptor inhibition modulates the nuclear localization and cytotoxicity of the Auger electron emitting radiopharmaceutical 111In-DTPA human epidermal growth factor. J Nucl Med. 2007 Sep;48(9):1562-70.
  18. Watanabe N, Sawai H, Ogihara-Umeda I, Tanada S, Kim EE, Yonekura Y, Sasaki Y. Molecular therapy of human neuroblastoma cells using Auger electrons of 111In-labeled N-myc antisense oligonucleotides. J Nucl Med. 2006 Oct;47(10):1670-7.
  19. Michel RB, Andrews PM, Castillo ME, Mattes MJ. In vitro cytotoxicity of carcinoma cells with 111In-labeled antibodies to HER-2. Mol Cancer Ther. 2005 Jun;4(6):927-37.
  20. Stokkel MP, Verkooijen RB, Bouwsma H, Smit JW. Six month follow-up after 111In-DTPA-octreotide therapy in patients with progressive radioiodine non-responsive thyroid cancer: a pilot study. Nucl Med Commun. 2004 Jul;25(7):683-90.
  21. Meyers MO, Anthony LB, McCarthy KE, Drouant G, Maloney TJ, Espanan GD, Woltering EA. High-dose indium 111In pentetreotide radiotherapy for metastatic atypical carcinoid tumor. South Med J. 2000 Aug;93(8):809-11.
  22. Collins AR. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol. 2004 Mar;26(3):249-61.
  23. Sadeghpour H, Jalilian AR, Akhlaghi M, Kamali-dehghan M, Mirzaii M. Preparation and biodistribution of [111In]-rHuEpo for erythropoietin receptor imaging. J Radioanal Nucl Chem. 2008;278:117-122.
  24. Alirezapour B, Jalilian AR1, Rajabifar S, Mirzaii M, Moradkhani S, Pouladi M, Aslani G. Preclinical evaluation of [¹¹¹ In]-DOTA-trastuzumab for clinical trials. J Cancer Res Ther. 2014 Jan-Mar;10(1):112-20.
  25. Pippin CG, Parker TA, McMurry TJ, Brechbiel MW. Spectrophotometric method for the determination of a bifunctional DTPA ligand in DTPA-monoclonal antibody conjugates. Bioconjug Chem. 1992 Jul-Aug;3(4):342-5.
  26. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680-5.
  27. Lindmo T, Boven E, Cuttitta F, Fedorko J, Bunn PA Jr. Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods. 1984 Aug 3;72(1):77-89.
  28. Matzku S, Moldenhauer G, Kalthoff H, Canevari S, Colnaghi M, Schuhmacher J, Bihl H. Antibody transport and internalization into tumours. Br J Cancer Suppl. 1990 Jul;10:1-5.
  29. Enciso M, Sarasa J, Agarwal A, Fernández JL, Gosálvez J. A two-tailed Comet assay for assessing DNA damage in spermatozoa.  Reprod Biomed Online. 2009 May;18(5):609-16.
  30. Frenzilli G, Bernardeschi M, Barale R. Alkaline versus neutral version of comet assay in human leukocytes using 9 compounds. J Transl Toxicol. 2014;1(1):60-71.
  31. Lewis MR, Boswell CA, Laforest R, Buettner TL, Ye D, Connett JM, Anderson CJ. Conjugation of monoclonal antibodies with TETA using activated esters: biological comparison of 64Cu-TETA-1A3 with 64Cu-BAT-2IT-1A3. Cancer Biother Radiopharm. 2001 Dec;16(6):483-94.
  32. Costantini DL, Chan C, Cai Z, Vallis KA, Reilly RM. (111)In-labeled trastuzumab (Herceptin) modified with nuclear localization sequences (NLS): an Auger electron-emitting radiotherapeutic agent for HER2/neu-amplified breast cancer. J Nucl Med. 2007 Aug;48(8):1357-68.
  33. Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol. 2001 May;Appendix 3:Appendix 3B.

Files

Share

How to cite

Statistics