Inhibition of papilloma progression by antisense oligonucleotides targeted to HPV11 E6/E7 RNA

G. A. Clawson, G. Q. Miranda, A. Sivarajah, P. Xin, W. Pan, D. Thiboutot, N. D. Christensen

Research output: Contribution to journalArticle

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Abstract

Human papillomaviruses (HPVs) are recognized as important human pathogens, causing a spectrum of hyperproliferative lesions from benign warts to cervical dysplasias/carcinomas. HPV-associated lesions require continued production of the oncogenic E6/E7 proteins, which are encoded by either bicistronic or overlapping mRNAs. Here we targeted the E6/E7 mRNA of HPV11, a type implicated in causation of genital warts, using molecular reagents. Accessible sites in the HPV11E6/E7 RNA were identified using library selection protocols, and nucleic acids (DNA-zymes, antisense oligonucleotides) targeted to these sites were constructed, and tested in cell culture and on human foreskin grafts. While DNAzymes were at least equally effective in cell culture, antisense oligonucleotides targeted to the region surrounding one of the library-selected sites (ASO407) proved most effective in blocking progression of HPV11-induced papillomas in human foreskin grafts on immunodeficient mice. In total, 11 papillomas were treated with ASO407. Of these, four of seven small papillomas treated with ASO407 showed loss of detectable virus by in situ hybridization (ISH), and in all four of these, papillomas were no longer evident grossly or histologically after treatment. When larger papillomas were treated, one of four showed loss of virus by ISH, associated with a minor decrease in papilloma size. Considering all 11 papillomas treated with ASO407 loss of viral staining by ISH was significantly different from that observed in controls (P < 0.016), as was true for the seven small treated papillomas (P < 0.012). DNAzymes targeted to the same site (or other library selected sites) did not produce statistically significant differences in ISH staining (P < 0.15). Our results with ASO417 appear to represent the first specific molecular therapy against a bona fide HPV infection, and provide a rational proof-of-principle strategy for development of molecular therapeutics targeting other HPV-associated lesions.

Original languageEnglish (US)
Pages (from-to)1331-1341
Number of pages11
JournalGene Therapy
Volume11
Issue number17
DOIs
StatePublished - Sep 1 2004

Fingerprint

Antisense Oligonucleotides
Papilloma
RNA
In Situ Hybridization
Catalytic DNA
Libraries
Foreskin
Cell Culture Techniques
Uterine Cervical Dysplasia
Staining and Labeling
Viruses
Transplants
Condylomata Acuminata
Messenger RNA
Warts
Papillomavirus Infections
Causality
Carcinoma
DNA
Therapeutics

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

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title = "Inhibition of papilloma progression by antisense oligonucleotides targeted to HPV11 E6/E7 RNA",
abstract = "Human papillomaviruses (HPVs) are recognized as important human pathogens, causing a spectrum of hyperproliferative lesions from benign warts to cervical dysplasias/carcinomas. HPV-associated lesions require continued production of the oncogenic E6/E7 proteins, which are encoded by either bicistronic or overlapping mRNAs. Here we targeted the E6/E7 mRNA of HPV11, a type implicated in causation of genital warts, using molecular reagents. Accessible sites in the HPV11E6/E7 RNA were identified using library selection protocols, and nucleic acids (DNA-zymes, antisense oligonucleotides) targeted to these sites were constructed, and tested in cell culture and on human foreskin grafts. While DNAzymes were at least equally effective in cell culture, antisense oligonucleotides targeted to the region surrounding one of the library-selected sites (ASO407) proved most effective in blocking progression of HPV11-induced papillomas in human foreskin grafts on immunodeficient mice. In total, 11 papillomas were treated with ASO407. Of these, four of seven small papillomas treated with ASO407 showed loss of detectable virus by in situ hybridization (ISH), and in all four of these, papillomas were no longer evident grossly or histologically after treatment. When larger papillomas were treated, one of four showed loss of virus by ISH, associated with a minor decrease in papilloma size. Considering all 11 papillomas treated with ASO407 loss of viral staining by ISH was significantly different from that observed in controls (P < 0.016), as was true for the seven small treated papillomas (P < 0.012). DNAzymes targeted to the same site (or other library selected sites) did not produce statistically significant differences in ISH staining (P < 0.15). Our results with ASO417 appear to represent the first specific molecular therapy against a bona fide HPV infection, and provide a rational proof-of-principle strategy for development of molecular therapeutics targeting other HPV-associated lesions.",
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Inhibition of papilloma progression by antisense oligonucleotides targeted to HPV11 E6/E7 RNA. / Clawson, G. A.; Miranda, G. Q.; Sivarajah, A.; Xin, P.; Pan, W.; Thiboutot, D.; Christensen, N. D.

In: Gene Therapy, Vol. 11, No. 17, 01.09.2004, p. 1331-1341.

Research output: Contribution to journalArticle

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AU - Clawson, G. A.

AU - Miranda, G. Q.

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