A novel drug screening assay for papillomavirus specific antiviral activity

Clark, Paul; Roberts, M L; Cowsert, Lex M.

doi:10.1016/s0166-3542(97)00066-1

Cited by 14 publications

(10 citation statements)

References 24 publications

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“…Although a study by others has shown only marginal efficacy of α-IFN in another culture system established with SCC-4 cells, this discrepancy may be due to the use of a different IFN-α and cell line. 9 In conclusion, we have developed a convenient and relatively stable in vitro cell model that may be used to screen anti-HPV-11 drugs. Our data have demonstrated that this system is easy to operate, and convenient to store and reproduce.…”

Section: Discussionmentioning

confidence: 99%

“…20 It is thought that the persistent and latent infections lead to an effective evasion of defensive system of the host cells, which results in the difficulty in thoroughly curing CA and the high recurrence rate after the initial remission following treatments. 9,21 Thus, the inhibition of the replication of HPV-11 DNA by anti-HPV agents at this stage may help the clearance of virus from the infected epithelial cells. To develop an effective in vitro screening system for anti–HPV-11 drugs, the critical step depends on the establishment of the replication and amplification of viral DNA in host cells.…”

Section: Discussionmentioning

confidence: 99%

“…To date, many attempts have been made to mimic HPV-11 infection in tissues in vitro. Several types of mammalian cells, such as SCC-4 cells (derived from a squamous cell carcinoma), primary human foreskin keratinocytes (HFKs), and immortalized human keratinocytes (N-Tert cells with the catalytic subunit of human telomerase and HaCaT cells), [9][10][11][12] have been used to establish HPV-11 DNA-containing cells by introducing gene segments, from the linear or circularized genome of HPV-11, with electroporation or chemical transfection techniques. In these cells, the replication of HPV-11 DNA in transient-or definite-passage cell cultures has been observed.…”

Section: Research-article2014mentioning

confidence: 99%

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Development of Basal-Like HaCaT Keratinocytes Containing the Genome of Human Papillomavirus (HPV) Type 11 for Screening of Anti-HPV Effects

Wang

Song

et al. 2014

SLAS Discovery

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Condylomata acuminata (CA), induced by low-risk human papillomaviruses (HPVs), is one of the most common sexually transmitted diseases. The increasing incidence and the high recurrence rate of CA have significantly contributed to public health problems around the world. Because HPVs cannot be cultured in vitro for a long time, there has been little progress in the development of HPV-specific antiviral agents. In this study, we established an HPV11.HaCaT system by introducing the recircularized genome of HPV-11 into HaCaT keratinocytes with transfection techniques and cultured them in a special medium. The existence and replication of HPV-11 DNA were positively detected in established HPV11.HaCaT cells. The HPV-11 DNA in HPV11.HaCaT cells has been stably replicated in definite passages of cells. We preliminarily studied the anti-HPV-11 effects of recombinant human interferon α1b (rhIFN-α) and 13-hexyl-palmatine hydrochloride (HP-13) in HPV11.HaCaT cells. The results suggest that HP-13 significantly inhibited the proliferation of HPV11.HaCaT cells in a dose-dependent manner, whereas rhIFN-α did not. HP-13 and rhIFN-α inhibited the replication of HPV-11 DNA and the expression of E1 ∧ E4 mRNA in HPV11.HaCaT cells. In conclusion, the established HPV11.HaCaT cells can provide us with a convenient and relatively stable tool for screening anti-HPV-11 agents.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Research-article2014mentioning

confidence: 99%

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Development of Basal-Like HaCaT Keratinocytes Containing the Genome of Human Papillomavirus (HPV) Type 11 for Screening of Anti-HPV Effects

Wang

Song

et al. 2014

SLAS Discovery

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“…Cowsert et al used phosphorothioate antisense oligonucleotides to block E2 mRNA translation and E2-mediated transcriptional trans-activation in cultured cells (7,9). More recently, a peptidic nucleic acid was designed to bind to a double-stranded DNA bovine papillomavirus E2-binding site, thereby antagonizing E2-DNA binding by peptide-DNA association.…”

Section: Vol 78 2004mentioning

confidence: 99%

“…Theoretically, a higher-affinity antagonist peptide would more fully inhibit E2-DNA binding in cell culture; however, it is also possible that the NLS and GFP regions of the fusion peptide caused some steric hindrance of 6N40D-E2 association. Note that transcription inhibition studies with phosphorothioate oligonucleotide E2-DNA binding antagonists (7) indicated that an oligonucleotide of lower E2-binding affinity caused only a 50% decrease in trans-activation activity, while an oligonucleotide of higher affinity decreased transactivation 10-fold. Thus, the incomplete suppression of E2-mediated trans-activation by the 6N40D-NLS-GFP fusion protein might simply be a reflection of suboptimal E2-DNA binding antagonism.…”

Section: Vol 78 2004mentioning

confidence: 99%

Identification of Peptides That Inhibit the DNA Binding,trans-Activator, and DNA Replication Functions of the Human Papillomavirus Type 11 E2 Protein

Deng¹,

Pearce²,

Dixon³

et al. 2004

J Virol

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Peptide antagonists of the human papillomavirus type 11 (HPV-11) E2-DNA association were identified using a filamentous bacteriophage random peptide library. Synthetic peptides antagonized the E2-DNA interaction, effectively blocked E2-mediated transcriptional activation of a reporter gene in cell culture, and inhibited E1-E2-mediated HPV-11 DNA replication in vitro. These peptides may prove to be useful tools for characterizing E2 function and for exploring the effectiveness of E2-inhibitor-based treatments for HPVassociated diseases.Present estimates are that 24 million Americans are infected with human papillomavirus (HPV) and that 5 million new cases occur annually. As there are no specific antiviral therapies for HPV infection, there is a clear unmet need for the development of safe and effective therapies for HPV-associated disease.Papillomaviruses lack the enzymes generally targeted by most currently available antiviral agents (e.g., virus-encoded DNA polymerases, reverse transcriptases, and proteases). Papillomaviruses do encode E1, a helicase-ATPase, but it has so far proven a difficult enzyme to develop for classic drug screening. As such, successful development of anti-HPV therapies may require identification of novel antiviral targets, such as viral transcription factors and replication proteins (2, 21, 25) such as the HPV-encoded E2 protein.The 50-kDa E2 protein is comprised of three functional domains (19). The amino-terminal domain of E2 is necessary for viral trans-activation and for direct association with E1. The smaller carboxyl-terminal domain (E2C) encodes the DNA binding and dimerization functions. Linking the amino-and carboxyl-terminal domains is a small, poorly conserved hinge region. E2 function requires binding to a 12-bp palindromic nucleotide sequence, ACCN 6 GGT, that is present at several locations throughout the HPV genome and is repeated several times near the viral origin of replication. Once associated with DNA, E2 interacts with a variety of host cell transcription factors to modulate viral transcription. The papillomavirus E2 protein is also required for origin (ORI)-specific viral DNA replication. Because association of E1 with E2 enhances the affinity of both proteins for the viral ORI, DNA sequencespecific binding by E2 effectively mediates sequence-specific DNA binding by E1 (6).Since the E2-DNA interaction is central to the coordination of the essential papillomavirus transcription and replication processes, disruption of E2-DNA binding should inhibit HPV replication. To this end, we constructed a filamentous bacteriophage 10-mer random peptide library and screened for sequences that disrupted the E2-DNA association by binding to the E2 protein.Phage display selection of E2-DNA binding inhibitors. We used peptide phage display (reviewed in reference 16) to identify peptide binders to bio-E2C, a biotinylated E2 fusion protein consisting of 88 carboxyl-terminal amino acids of the HPV type 11 (HPV-11) E2 DNA binding domain fused to the 99-residue polypeptide substrate of Es...

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Protein profiling and identification of modulators regulated by the E7 oncogene in the C33A cell line by proteomics and genomics

Lee¹,

Shim²,

Kho

et al. 2004

Proteomics

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Human papillomaviruses (HPVs) have been recognized as the primary cause of cervical cancer. HPV 16 E7 binds to tumor suppressor retinoblastoma protein, and interferes with its function, causing release of the transcription factor E2F, which influences expression of cell cycle-related genes. This study was performed to identify the genes and proteins modulated by the HPV E7 oncogene. An HPV-negative cervical cancer cell line (C33A) was prepared to establish a stable cell line expressing E7. In order to analyze the target molecules modulated by E7 expression, we used two approaches: matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and DNA microarrays. Forty-seven spots were identified in C33A/E7 by two-dimensional electrophoresis and MALDI/TOF MS. Protein disulfide isomerase A3, integrase interactor 1 protein, growth inhibitory protein, glutathione S-transferase P, and vav proto-oncogene were down-regulated, whereas heat shock 60 kDa protein 1, Ku70 binding protein, alpha enolase, 26S proteasome subunit were up-regulated. A genomic approach using a microarray kit showed that IL-12R beta 1, cytochrome c, and tumor necrosis factor receptor II were induced by the E7 oncogene. These results suggest that E7 can evade immune surveillance by suppressing or inducing these cell signaling factors, cell cycle regulators, and chaperones.

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A novel drug screening assay for papillomavirus specific antiviral activity

Cited by 14 publications

References 24 publications

Development of Basal-Like HaCaT Keratinocytes Containing the Genome of Human Papillomavirus (HPV) Type 11 for Screening of Anti-HPV Effects

Development of Basal-Like HaCaT Keratinocytes Containing the Genome of Human Papillomavirus (HPV) Type 11 for Screening of Anti-HPV Effects

Identification of Peptides That Inhibit the DNA Binding,trans-Activator, and DNA Replication Functions of the Human Papillomavirus Type 11 E2 Protein

Protein profiling and identification of modulators regulated by the E7 oncogene in the C33A cell line by proteomics and genomics

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