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Inhibition of the human chemokine receptor CXCR4 by antisense phosphorothioate oligodeoxyribonucleotides
A Kusunoki1, T Saitou, N Miyano-Kurosaki
1Department of Industrial Chemistry, Chiba Institute of Technology, Narashino, Chiba, Japan.
Abstract:
The CXC chemokine receptor CXCR4/fusion, a major coreceptor for the T-cell line T-tropic (X4) HIV-1 virus, plays a critical role in T-tropic virus fusion and entry into permissive cells. In the present study, we describe the effects of an antisense phosphorothioate oligodeoxyribonucleotide (anti-S-ODN) on the inhibition of CXCR4 gene expression in X4 HIV-1 infected HeLa-CD4 cells, to find more efficacious therapeutic possibilities for human immunodeficiency virus type 1 (HIV-1) infection. The naked antisense phosphorothioate oligodeoxyribonucleotide (anti-S-ODN-1), containing the AUG initiation codon at the center of the oligodeoxyribonucleotide, showed a slightly higher inhibitory effect on HIV-1 gag p24 production among all sequences tested. We also examined the concomitant use of a basic peptide transfection reagent, nucleosomal histone proteins (RNP), for the delivery of the anti-S-ODN-1. The anti-S-ODN-1 encapsulated with RNP had higher inhibitory effects on p24 products than the naked anti-S-ODN-1. When the anti-S-ODN-1 encapsulated with RNP was incubated with HeLa-CD4 cells, the surface levels of this chemokine receptor showed high suppression, indicating sequence-specific inhibition. The activities of unmodified oligodeoxyribonucleotide are effectively enhanced by using a basic peptide, RNP.
Insights
An antisense oligodeoxynucleotide targeting CXCR4 gene expression inhibited HIV-1 replication. Delivery with RNP enhanced this inhibition, offering a potential therapeutic strategy for HIV-1 infection.
Area of Science:
- Virology
- Molecular Biology
- Drug Discovery
Background:
- CXC chemokine receptor CXCR4 is essential for T-tropic (X4) HIV-1 entry.
- HIV-1 infection remains a significant global health challenge.
- Targeting viral entry pathways is a key therapeutic strategy.
Purpose of the Study:
- To investigate the efficacy of an antisense phosphorothioate oligodeoxyribonucleotide (anti-S-ODN) in inhibiting CXCR4 gene expression.
- To evaluate the potential of anti-S-ODN as a therapeutic agent against X4 HIV-1 infection.
- To assess the role of nucleosomal histone proteins (RNP) in enhancing anti-S-ODN delivery and efficacy.
Main Methods:
- Synthesized and tested various anti-S-ODN sequences for their inhibitory effects on HIV-1 gag p24 production in HeLa-CD4 cells.
- Utilized a basic peptide transfection reagent, RNP, for the delivery of anti-S-ODN.
- Quantified the suppression of CXCR4 surface levels on treated cells to confirm sequence-specific inhibition.
Main Results:
- The anti-S-ODN-1 sequence demonstrated a notable inhibitory effect on HIV-1 gag p24 production.
- Encapsulation of anti-S-ODN-1 with RNP significantly enhanced its inhibitory activity compared to naked anti-S-ODN-1.
- Incubation with RNP-encapsulated anti-S-ODN-1 led to substantial suppression of CXCR4 surface expression, confirming sequence-specific gene silencing.
Conclusions:
- Antisense oligodeoxyribonucleotides targeting CXCR4 show promise for inhibiting HIV-1 replication.
- RNP-mediated delivery enhances the efficacy of antisense oligodeoxyribonucleotides.
- This approach presents a potential therapeutic avenue for managing HIV-1 infection by blocking viral entry.