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Antisense phosphorothioate oligonucleotides targeted to the human chemokine receptor CXCR4
A Kusunoki1, N Miyano-Kurosaki, T Kimura
1Department of Industrial Chemistry, Chiba Institute of Technology, Narashino, Japan.
Abstract:
The CXC chemokine receptor CXCR4 is used as a major co-receptor for fusion and entry by syncytia-inducing T-tropic (X4) isolates of HIV-1. In the present study, we report the effects of an antisense oligodeoxyribonucleotide 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. Antisense phosphorothioate oligodeoxyribonucleotides (anti-S-ODNs) corresponding to the sequence of bases 69 to 88 of the human CXCR4 mRNA gene were synthesized. When the naked anti-S-ODN was incubated with HeLa-CD4 cells, the surface levels of this chemokine receptor were reduced up to 50%, indicating sequence-specific inhibition. We also examined the concomitant use of a basic peptide transfection reagent, nucleosomal histone proteins (RNP), for delivery of anti-S-ODNs. The anti-S-ODN encapsulated with RNP had higher inhibitory effects on p24 products than the naked anti-S-ODN.
Insights
Antisense oligodeoxyribonucleotides targeting the CXCR4 gene effectively reduced its expression in Human Immunodeficiency Virus type 1 (HIV-1) infected cells. Delivery using RNP enhanced inhibition of viral products, suggesting a potential therapeutic strategy.
Area of Science:
- Molecular Biology
- Virology
- Immunology
Background:
- CXC chemokine receptor CXCR4 is a critical co-receptor for Human Immunodeficiency Virus type 1 (HIV-1) entry.
- Syncytia-inducing T-tropic (X4) HIV-1 isolates heavily rely on CXCR4 for cell fusion and infection.
Purpose of the Study:
- To investigate the therapeutic potential of inhibiting CXCR4 gene expression using antisense oligodeoxyribonucleotides (anti-S-ODNs).
- To evaluate the efficacy of anti-S-ODNs in X4 HIV-1 infected cells.
Main Methods:
- Synthesis of antisense phosphorothioate oligodeoxyribonucleotides (anti-S-ODNs) targeting human CXCR4 mRNA.
- Treatment of HeLa-CD4 cells infected with X4 HIV-1 using naked anti-S-ODNs.
- Assessment of CXCR4 surface receptor levels via flow cytometry.
- Evaluation of anti-S-ODN delivery enhancement using nucleosomal histone proteins (RNP).
Main Results:
- Naked anti-S-ODNs reduced surface CXCR4 levels by up to 50% in a sequence-specific manner.
- Anti-S-ODNs encapsulated with RNP demonstrated superior inhibition of p24 viral products compared to naked anti-S-ODNs.
Conclusions:
- Antisense oligodeoxyribonucleotides are effective in inhibiting CXCR4 gene expression in X4 HIV-1 infected cells.
- RNP-mediated delivery of anti-S-ODNs enhances therapeutic efficacy against HIV-1, offering a promising avenue for treatment.