Searchable abstracts of presentations at key conferences in endocrinology
Endocrine Abstracts (2007) 13 P142

SFEBES2007 Poster Presentations Diabetes, metabolism and cardiovascular (63 abstracts)

Antisense oligonucleotide-induced alternative splicing of the APOB mRNA generates a novel isoform of APOB

Bernard Khoo 1 , Xavier Roca 2 , Shern L Chew 1 & Adrian R Krainer 2


1Queen Mary University of London, London, United Kingdom; 2Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States.


Background: Apolipoprotein B (APOB) is an integral part of the LDL, VLDL, IDL, Lp(a) and chylomicron lipoprotein particles. The APOB pre-mRNA consists of 29 constitutively-spliced exons. APOB exists as two natural isoforms: the full-length APOB100 isoform, assembled into LDL, VLDL, IDL and Lp(a) and secreted by the liver in humans; and the C-terminal truncation APOB48, assembled into chylomicrons and secreted by the intestine in humans. Down-regulation of APOB100 is a potential therapy to lower circulating LDL and cholesterol levels.

Results: We investigated the ability of 2′O-methyl RNA antisense oligonucleotides (ASOs) to induce skipping of exon 27 in endogenous APOB mRNA in HepG2 cells. These ASOs are directed towards the 5′ and 3′ splice-sites of exon 27, the branch point sequence (BPS) of intron 26–27 and several predicted splicing enhancers within exon 27. ASOs targeting either the 5′ or 3′ splice-site, in combination with the BPS, are the most effective. The splicing of other alternatively spliced genes are not influenced by these ASOs, suggesting that the effects are not due to non-specific changes in alternative splicing. The skip 27 mRNA is translated into a truncated isoform, APOB87SKIP27.

Conclusions: Patients with hypobetalipoproteinaemia have mutations in APOB that cause the expression of C-terminal truncations of APOB. LDL, VLDL and IDL containing these truncations are more rapidly cleared, as the truncated APOB has a higher affinity for the LDL receptor. We predict the induction of APOB87SKIP27 expression in vivo should lead to decreased LDL and cholesterol levels. As intestinal APOB mRNA editing and APOB48 expression rely on sequences within exon 26, exon 27 skipping should not affect APOB48 expression unlike other methods of down-regulating APOB100, such as RNA interference, which are known to also down-regulate APOB48. This technique represents a promising therapeutic method to reduce LDL and cholesterol levels in vivo.

Article tools

My recent searches

No recent searches.