Targeting a Pathogenic Variant Creating an Upstream AUG in the ENG 5' Untranslated Region with Antisense Oligonucleotides Fails to Restore Protein Expression
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View abstract on PubMed
Summary
This summary is machine-generated.Antisense oligonucleotides (ASOs) failed to restore endoglin protein levels in hereditary hemorrhagic telangiectasia (HHT) models caused by 5'UTR variants. Further research is needed to understand uORF-mediated translation regulation.
Area Of Science
- Genetics
- Molecular Biology
- Biochemistry
Background
- Pathogenic variants in the 5' untranslated region (5'UTR) of the ENG gene, specifically those creating upstream open reading frames (uORFs), can disrupt translation and lead to hereditary hemorrhagic telangiectasia (HHT).
- The ENG c.-79C>T variant introduces a uAUG that reduces endoglin expression, contributing to HHT.
Purpose Of The Study
- To investigate the potential of 2'-O-methyl (2'OMe) antisense oligonucleotides (ASOs) to restore endoglin protein levels.
- To assess if ASOs can mask the aberrant uAUG or target secondary structures in the ENG 5'UTR.
Main Methods
- ASOs of varying lengths and chemistries were designed to target the mutant region of the ENG 5'UTR.
- Experiments were conducted in HeLa cells and human umbilical vein endothelial cells (HUVECs) transfected/transduced with wild-type or mutant ENG constructs.
- Transfection efficiency was confirmed by MALAT1 knockdown via qPCR, and endoglin levels were measured by Western blot.
Main Results
- Despite successful ASO delivery and optimized conditions, no consistent increase in endoglin protein levels was observed after ASO treatment.
- The study demonstrated limitations of steric-blocking ASOs for targeting 5'UTR variants that cause uORF-mediated translation disruption.
Conclusions
- Steric-blocking ASOs may not be effective in restoring protein levels for all 5'UTR variants causing HHT.
- Further investigation into the complex mechanisms of uORF-mediated translational regulation is essential.
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