Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: Anti-proliferative effects on HPV 16-transformed cells

Francesca Venturini; Joris Braspenning; Matthias Homann; Lutz Gissmann; Georg Sczakiel

doi:10.1093/nar/27.7.1585

Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: Anti-proliferative effects on HPV 16-transformed cells

Francesca Venturini, Joris Braspenning, Matthias Homann, Lutz Gissmann, Georg Sczakiel^*

^*Corresponding author for this work

Institute of Molecular Medicine

German Cancer Research Center

57 Citations (Scopus)

Abstract

The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 10⁴ M^-1s^-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.

Original language	English
Journal	Nucleic Acids Research
Volume	27
Issue number	7
Pages (from-to)	1585-1592
Number of pages	8
ISSN	0305-1048
DOIs	https://doi.org/10.1093/nar/27.7.1585
Publication status	Published - 01.04.1999

Funding

We thank O. Wildner for initial support as well as A. Hörster and R. Kronenwett for help with FACS analysis and transfection experiments. We also thank B. Teichmann and V. Patzel for critical comments on the manuscript. This work was supported by the Bundesministerium für Bildung, Forschung und Wissenschaft (grant no. 01KV9517) and the Fonds der Chemischen Industrie.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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title = "Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: Anti-proliferative effects on HPV 16-transformed cells",

abstract = "The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 104 M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.",

author = "Francesca Venturini and Joris Braspenning and Matthias Homann and Lutz Gissmann and Georg Sczakiel",

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AU - Gissmann, Lutz

AU - Sczakiel, Georg

N1 - Funding Information: We thank O. Wildner for initial support as well as A. Hörster and R. Kronenwett for help with FACS analysis and transfection experiments. We also thank B. Teichmann and V. Patzel for critical comments on the manuscript. This work was supported by the Bundesministerium für Bildung, Forschung und Wissenschaft (grant no. 01KV9517) and the Fonds der Chemischen Industrie.

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N2 - The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 104 M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.

AB - The E6/E7-coding sequences of the human papillomavirus type 16 (HPV 16) were probed for kinetic accessibility in vitro by pools of catalytic antisense RNA. Only long-chain complementary RNA and very few antisense sequences with a 3' portion complementary to a 10 nt window within unspliced and spliced E6-coding target sequences showed fast annealing with k(ass) values of up to 104 M-1s-1 indicating that the majority of E6/E7 RNA sequences are unfavourable targets for antisense inhibitors and ribozymes. Fast-annealing antisense oligodeoxyribonucleotides directed against the window of 10 nt inhibited cell proliferation of HPV 16-transformed SiHa cells but not slow-annealing antisense species. Antisense RNA of several hundred nucleotides in length also showed significant antiproliferative activity. Biological effects of antisense oligodeoxyribonucleotides were specific for the antisense sequence, could only be found in HPV-positive but not in HPV-negative cell lines, and were related to decreased levels of E7 protein and E6/E7-specific transcripts. This work suggests that HPV 16 E7/E6 sequences exhibit a low accessibility for antisense oligonucleotides. This can be overcome, however, by exploiting the relationship between fast annealing of antisense species and their increased efficacy in human cells.

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Kinetic selection of HPV 16 E6/E7-directed antisense nucleic acids: Anti-proliferative effects on HPV 16-transformed cells

Abstract

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