Inhibition of Caspase-2 Activity in Human Jurkat T-cell Lymphoma Cells by Splice Switching Oligonucleotide to its pre-mRNA

  • D.D. Zhdanov Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia; Peoples Friendship University of Russia, 6 Miklukho-Maklaya str., Moscow, 117198 Russia
  • A.A. Plyasova Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
  • Yu.A. Gladilina Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
  • M.V. Pokrovskaya Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
  • S.S. Alexandrova Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
  • N.N. Sokolov Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
Keywords: caspase-2; alternative splicing; splice switching oligonucleotide; enzymatic activity


Caspase-2 is a key enzyme thinvolved in induction of apoptosis. The caspase-2 level is regulated by alternative splicing (AS) of its mRNA. The aim of this work was to determine the ability of an oligonucleotide complementary to Casp-2 pre-mRNA to induce AS. This oligonucleotide blocked the binding of splicing-regulating proteins to their sites at the end of exon 9 of Casp-2 pre-mRNA, leading to induction of AS of Casp-2 mRNA. The decrease in expression of full-size active splice-variant (Casp-2L) and the increase the expression of a shortened variant (Casp-2S) was demonstrated in human T-cell lymphoma Jurkat cell line. The expression level of total Casp-2 remained unchanged. Disproportion of splice variants of Casp-2 led to inhibition of enzymatic activity of caspase-2.


  1. Bao, Q., Shi, Y. (2007) Apoptosome: a platform for the activation of initiator caspases. Cell Death and Differentiation, 14(1), 56–65. DOI

  2. Vakifahmetoglu-Norberg, H., Zhivotovsky, B. (2010) The unpredictable caspase-2: what can it do? Trends in Cell Biology, 20(3), 150–159. DOI

  3. Wotawa, A., Solier, S., Logette, E., Solary, E., Corcos, L. (2002) Differential influence of etoposide on two caspase-2 mRNA isoforms in leukemic cells. Cancer Letters, 185(2), 181–189. DOI

  4. Aravind, L., Dixit, V.M., Koonin, E.V. (1999) The domains of death: evolution of the apoptosis machinery. Trends in Biochemical Sciences, 24(2), 47–53. DOI

  5. Zhivotovsky, B., Orrenius, S. (2005) Caspase-2 function in response to DNA damage. Biochemical and Biophysical Research Communications, 331(3), 859–867. DOI

  6. Wang, L., Miura, M., Bergeron, L., Zhu, H., Yuan, J. (1994) Ich-1, an Ice/ced-3-related gene, encodes both positive and negative regulators of programmed cell death. Cell, 78(5), 739–750. DOI

  7. Kumar, S., Kinoshita, M., Noda, M. (1997) Characterization of a mammalian cell death gene Nedd2. Leukemia, 11(3), 385–386. DOI

  8. Lavrik, I.N., Golks, A., Baumann, S., Krammer, P.H. (2006) Caspase-2 is activated at the CD95 death-inducing signaling complex in the course of CD95-induced apoptosis. Blood, 108(2), 559–565. DOI

  9. Jiang, Z.H., Zhang, W.J., Rao, Y., Wu, J.Y. (1998) Regulation of Ich-1 pre-mRNA alternative splicing and apoptosis by mammalian splicing factors. Proc. Natl. Acad. Sci. USA, 95(16), 9155–9160. DOI

  10. Côté, J., Dupuis, S., Wu, J.Y. (2001) Polypyrimidine track-binding protein binding downstream of caspase-2 alternative exon 9 represses its inclusion. Journal of Biological Chemistry, 276(11), 8535–8543. DOI

  11. Havlioglu, N., Wang, J., Fushimi, K., Vibranovski, M.D., Kan, Z., Gish, W., Wu, J.Y. (2007) An intronic signal for alternative splicing in the human genome. PloS one, 2(11), e1246. DOI

  12. Fushimi, K., Ray, P., Kar, A., Wang, L., Sutherland, L.C., Wu, J.Y. (2008) Up-regulation of the proapoptotic caspase 2 splicing isoform by a candidate tumor suppressor, RBM5. Proc. Natl. Acad. Sci. USA, 105(41), 15708–15713. DOI

  13. Rocha, C.S.J. (2019) Antisense Oligonucleotides for Splice Modulation: Assessing Splice Switching Efficacy. Methods Mol. Biol. 2036, 73–90. DOI

  14. Zhdanov, D.D., Gladilina, Y.A., Grishin, D.V., Pokrovsky, V.S., Pokrovskaya, M.V., Aleksandrova, S. S., Sokolov, N. N. (2018) Apoptotic Endonuclease EndoG Induces Alternative Splicing of Telomerase TERT Catalytic Subunit, Caspase-2, DNase I, and BCL-x in Human, Murine, and Rat CD4+T Lymphocytes. Russian Journal of Bioorganic Chemistry, 44(1), 90–103. DOI

  15. Zhdanov, D.D., Vasina, D.A., Orlova, V.S., Gotovtseva, V.Y., Bibikova, M. V., Pokrovsky, V.S., Sokolov, N.N. (2016) Apoptotic endonuclease EndoG induces alternative splicing of telomerase catalytic subunit hTERT and death of tumor cells. Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry, 10(4), 310–321. DOI

  16. Zhdanov, D.D., Gladilina, Y.A., Pokrovsky, V.S., Grishin, D.V., Grachev, V.A., Orlova, V.S., Sokolov, N.N. (2019) Endonuclease G modulates the alternative splicing of deoxyribonuclease 1 mRNA in human CD4+ T lymphocytes and prevents the progression of apoptosis. Biochimie, 157, 158–176. DOI

  17. Baker, B.F., Lot, S.S., Condon, T.P., Cheng-Flournoy, S., Lesnik, E.A., Sasmor, H.M., Bennett, C. F. (1997) 2’-O-(2-Methoxy)ethyl-modified anti-intercellular adhesion molecule 1 (ICAM-1) oligonucleotides selectively increase the ICAM-1 mRNA level and inhibit formation of the ICAM-1 translation initiation complex in human umbilical vein endothelial cells. Journal of Biological Chemistry, 272(18), 11994–12000. DOI

  18. Rigo, F., Seth, P.P., Bennett, C.F. (2014) Antisense oligonucleotide-based therapies for diseases caused by pre-mRNA processing defects. Advances in Experimental Medicine and Biology, 825, 303–352. DOI

  19. Havens, M.A., Hastings, M.L. (2016) Splice-switching antisense oligonucleotides as therapeutic drugs. Nucleic Acids Research, 44(14), 6549–6563. DOI