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Published on: May 11, 2018
Splice-Switching Antisense Oligonucleotides Targeting Extra- and Intracellular Domains of Epidermal Growth Factor
Akilandeswari Ashwini Balachandran1,2, Prithi Raguraman1,2, Kamal Rahimizadeh1,2
1Centre for Molecular Medicine and Innovative Therapeutics, Murdoch University, Murdoch, WA 6150, Australia.
Abstract:
Cancer is one of the leading causes of death globally. Epidermal growth factor receptor is one of the proteins involved in cancer cell proliferation, differentiation, and invasion. Antisense oligonucleotides are chemical nucleic acids that bind to target messenger ribonucleic acid and modulate its expression. Herein, we demonstrate the efficacy of splice-modulating antisense oligonucleotides to target specific exons in the extracellular (exon 3) and intracellular (exon 18, 21) domains of epidermal growth factor receptor. These antisense oligonucleotides were synthesized as 25mer 2'-O methyl phosphorothioate-modified ribonucleic acids that bind to complementary specific regions in respective exons. We found that PNAT524, PNAT525, PNAT576, and PNAT578 effectively skipped exon 3, exon 18, and exon 21 in glioblastoma, liver cancer, and breast cancer cell lines. PNAT578 treatment also skipped partial exon 19, complete exon 20, and partial exon 21 in addition to complete exon 21 skipping. We also found that a cocktail of PNAT576 and PNAT578 antisense oligonucleotides performed better than their individual counterparts. The migration potential of glioblastoma cancer cells was reduced to a greater extent after treatment with these antisense oligonucleotides. We firmly believe that using these splice-modulating antisense oligonucleotides in combination with existing EGFR-targeted therapies could improve therapeutic outcomes.
Insights
New antisense oligonucleotides effectively target and skip specific exons in epidermal growth factor receptor (EGFR), reducing cancer cell migration. Combining these splice-modulating oligonucleotides shows enhanced efficacy for potential cancer therapies.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Cancer remains a leading global cause of death.
- Epidermal growth factor receptor (EGFR) plays a crucial role in cancer cell progression.
- Antisense oligonucleotides offer a method to modulate gene expression by targeting messenger RNA.
Purpose of the Study:
- To investigate the efficacy of splice-modulating antisense oligonucleotides targeting specific exons of EGFR.
- To evaluate the impact of these oligonucleotides on cancer cell behavior, specifically migration.
Main Methods:
- Synthesis of 25mer 2'-O methyl phosphorothioate-modified ribonucleic acid antisense oligonucleotides.
- Treatment of glioblastoma, liver, and breast cancer cell lines with specific antisense oligonucleotides (PNAT524, PNAT525, PNAT576, PNAT578).
- Assessment of exon skipping and cell migration potential post-treatment.
Main Results:
- PNAT524, PNAT525, PNAT576, and PNAT578 successfully induced skipping of EGFR exons 3, 18, and 21 in various cancer cell lines.
- PNAT578 demonstrated broader exon skipping, including partial skipping of exon 19 and complete skipping of exon 20.
- A combination of PNAT576 and PNAT578 exhibited superior efficacy compared to individual treatments.
- Significant reduction in glioblastoma cell migration was observed following treatment.
Conclusions:
- Splice-modulating antisense oligonucleotides are effective in targeting specific EGFR exons.
- Combined antisense oligonucleotide therapy demonstrates enhanced anti-cancer effects, particularly on cell migration.
- These findings suggest a promising strategy for combining novel antisense oligonucleotides with existing EGFR-targeted therapies to improve cancer treatment outcomes.
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