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Liposomal targeting of bcl-2 antisense oligonucleotides with enhanced stability into human myeloma cell lines
H Ollikainen1, K Lappalainen, I Jääskeläinen
1MediCity Research Laboratory, University of Turku, Finland.
Abstract:
Cationic liposomes improve the delivery of antisense oligonucleotides (ODNs) into cells. However, there is marked variability in the cellular uptake of ODNs into different cell lines. We used liposomes containing dimethyloctadecylammonium bromide (DDAB) and dioleoylphosphatidylethanolamine (DOPE) to increase the delivery of phosphodiester ODNs into four different myeloma cell lines. The delivery by cationic liposomes increased the delivery of bcl-2 antisense ODNs by a factor of 9 to 45 as compared to plain ODNs. The stability of ODNs was increased with liposomes both in the culture medium and within the cells. Intact liposomal ODNs were detected inside the cells up to 24 hours with gel electrophoresis and phosphor imager analysis. Antisense ODNs had no effect on bcl-2 mRNA levels. Also the proliferation of myeloma cells remained unchanged during the 3-day incubation period. Our study shows that liposomal antisense ODNs targeting bcl-2 of human myeloma cells result in increased stability of ODNs with minimal toxicity. However, further modifications are needed to gain biological effects of antisense ODNs on human myeloma cells.
Insights
Cationic liposomes enhance the cellular delivery and stability of antisense oligonucleotides (ODNs) in myeloma cells. However, this method did not affect bcl-2 mRNA levels or cell proliferation, indicating a need for further optimization.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Cationic liposomes enhance antisense oligonucleotide (ODN) delivery into cells.
- Cellular uptake of ODNs varies significantly across different cell lines.
- Targeting bcl-2 in myeloma cells is a potential therapeutic strategy.
Purpose of the Study:
- To evaluate the efficacy of cationic liposomes (DDAB/DOPE) in delivering phosphodiester ODNs into myeloma cell lines.
- To assess the impact of liposomal delivery on ODN stability and cellular uptake.
- To determine the biological effects of bcl-2 antisense ODNs on myeloma cell proliferation and gene expression.
Main Methods:
- Liposomes composed of dimethyloctadecylammonium bromide (DDAB) and dioleoylphosphatidylethanolamine (DOPE) were formulated.
- Delivery of bcl-2 antisense ODNs using these liposomes into four myeloma cell lines was performed.
- Cellular uptake, ODN stability in vitro and in vivo, bcl-2 mRNA levels, and cell proliferation were analyzed.
Main Results:
- Liposomal delivery increased bcl-2 antisense ODN delivery by 9- to 45-fold compared to free ODNs.
- ODN stability was enhanced in culture medium and within cells, with intact liposomal ODNs detected up to 24 hours.
- No significant changes in bcl-2 mRNA levels or myeloma cell proliferation were observed.
Conclusions:
- Liposomal formulation enhances the stability and cellular delivery of antisense ODNs targeting bcl-2 in human myeloma cells with minimal toxicity.
- Further modifications are required to achieve a biological effect on bcl-2 mRNA levels and cell proliferation in myeloma.
- This study provides a foundation for developing improved liposomal antisense oligonucleotide strategies for cancer therapy.