Lowering LDL cholesterol levels by antisense oligonucleotide induced alternative splicing of Apolipoprotein B

Lead Research Organisation: University College London
Department Name: Unlisted


People with high levels of cholesterol are at higher risk of having heart attacks and strokes. Treatments that lower the levels of cholesterol have been shown to protect against these diseases, thereby saving lives. This project will look into how we can use a new drug called an antisense oligonucleotide to lower the level of cholesterol in the blood and therefore perhaps to reduce the risk of heart attacks and strokes.

Technical Summary

Hypercholesterolaemia and high LDL levels cause cardiovascular disease, the major killer in the developed world. Currently licensed treatments for hypercholesterolaemia have many shortcomings: there is a need for alternative therapies to lower cholesterol. Altering the expression of Apolipoprotein B (APOB) has emerged as a key method to accomplish this goal. APOB is the principal structural apolipoprotein in LDL, VLDL, IDL and chylomicron particles. Antisense oligonucleotides (ASOs) which induce alternative splicing of APOB exon 27 are able to generate a truncated APOB isoform, APOB87(SKIP27). Truncation of APOB100, as seen in patients with hypobetalipoproteinaemia, causes major reductions in total and LDL cholesterol levels. ASO-induced alternative splicing will be a useful therapy to reduce circulating LDL and cholesterol levels, without the drawbacks associated with methods that generally down-regulate all isoforms of APOB. In this project, we propose to: (1) optimize the configuration of ASOs and the skipping effect in vitro; and (2) assess the effects of injected optimized ASO on circulating LDL and cholesterol levels in a transgenic mouse model of hypercholesterolaemia and atherosclerosis. Completion of these objectives would validate this ASO-based compound as a therapy for hypercholesterolaemia.


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Disterer P (2014) Development of therapeutic splice-switching oligonucleotides. in Human gene therapy

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Disterer P (2012) Antisense-mediated exon-skipping to induce gene knockdown. in Methods in molecular biology (Clifton, N.J.)

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Disterer P (2013) Exon skipping of hepatic APOB pre-mRNA with splice-switching oligonucleotides reduces LDL cholesterol in vivo. in Molecular therapy : the journal of the American Society of Gene Therapy

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Khoo B (2015) Genetic therapies to lower cholesterol. in Vascular pharmacology

Description Proof-of-Concept Funding
Amount £25,205 (GBP)
Organisation UCL Business 
Sector Private
Country United Kingdom
Start 08/2012 
End 02/2013
Description Proof-of-Concept Funding
Amount £41,017 (GBP)
Organisation UCL Business 
Sector Private
Country United Kingdom
Start 11/2011 
End 05/2012
Description COST Network for Exon Skipping 
Organisation European Cooperation in Science and Technology (COST)
Country Belgium 
Sector Public 
PI Contribution Dr Bernard Khoo and Dr Petra Disterer have been elected to the COST Action BM1207 (for exon skipping)
Collaborator Contribution THis network allows access to partner expertise on topics relating to therapeutic development of antisense oligonucleotides for exon skipping
Impact We have made contact with potential collaborators regarding SSO deliver.
Start Year 2013
Description Collaboration with Acuitas Tx 
Organisation Acuitas Therapeutics
Country Canada 
Sector Private 
PI Contribution We are providing APO-skip SSOs for formulation testing and will be testing the formulations in human APOB mice.
Collaborator Contribution Acuitas is formulating the APO-skip SSOs in their lipid nanoparticles (previously used in human trials).
Impact Project is still on-going
Start Year 2013
Description Delivery of Antisense Oligonucleotides to Liver 
Organisation Life Technologies
Country Global 
Sector Private 
PI Contribution We are supplying the splice-switching oligonucleotides and testing these in transgenic mice.
Collaborator Contribution Life Technologies are developing new formulations for delivery of antisense oligonucleotides. Contribution: new Invivofectamine formulations.
Impact Experiments in progress
Start Year 2011
Description Methods for detection of oligonucleotides and APOB isoforms 
Organisation Quotient Bioresearch
Department Bioanalytical Sciences
Country United Kingdom 
Sector Private 
PI Contribution We are supplying materials (oligonucleotides, plasma samples)
Collaborator Contribution Development of mass-spectrometry methods to detect oligonucleotides and APOB isoforms
Impact Experiments in progress
Start Year 2012
Title Truncation of Apolipoprotein B100 with antisense oligonucleotides 
Description The invention concerns the use of an antisense oligonucleotide (ASO) to induce alternative splicing of Apolipoprotein B (APOB) by skipping exon 27. This is intended to truncate APOB100 expression and to lower LDL cholesterol for therapeutic purposes. INVENTIONS PATENTED 1. The concept of 'triple-targeting' three splice elements with a single ASO or combinations of ASOs (5' splice site, 3' splice site, branchpoint sequence) to increase exon skipping efficiency. 2. A new antisense oligonucleotide sequence for efficiently inducing APOB exon 27 skipping: s27_53B4. 3. The use of 5' fluoresceination of ASOs to increase exon skipping efficiency of ASOs. 4. The use of partial phosphorothioate backbone modification to increase exon skipping efficiency of ASOs. 5. The combination of any, or all of these modifications to increase exon skipping efficiency of ASOs. 
IP Reference GB1117880.3 
Protection Patent application published
Year Protection Granted 2011
Licensed No
Impact N/A
Title Antisense oligonucleotide to cause APOB exon 27 skipping 
Description Pre-clinical lead identified from cellular assays. In testing in animals. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Refinement. Non-clinical
Year Development Stage Completed 2010
Development Status Actively seeking support
Impact New technologies relating to modification of antisense oligonucleotides to increase efficiency in exon skipping identified.