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RNAi-mediated knockdown of HMG CoA reductase enhances gene expression from physiologically regulated low-density lipoprotein receptor therapeutic vectors in vivo

Gene Therapy volume 19, pages 463–467 (2012)Cite this article

Abstract

The development of novel strategies to enhance gene expression from therapeutic vectors may prove advantageous for complementation gene therapy. This applies to therapeutic expression of the low-density lipoprotein receptor (LDLR) gene to treat familial hypercholesterolaemia (FH), where appropriate gene regulation could enhance therapeutic effect. We have previously reported that LDLR genomic DNA expression vectors can be regulated in vivo by pravastatin. In the current study, we investigated whether targeted knockdown of the mevalonate pathway in conjunction with LDLR delivery would lead to enhanced LDLR transgene expression and improved phenotype recovery. We demonstrated here that knockdown of HMG CoA reductase (HMGCR) by up to 70% using small interfering RNAs (siRNAs) led to a significant increase in binding and internalisation of LDL particles in vitro in mouse and human cells. In vivo co-injection of LDLR promoter luciferase expression plasmids with siRNAs or microRNA (miRNA) expression vectors targeting mouse Hmgcr led to at least a 10-fold increase in luciferase expression. Injection of Ldlr−/− mice with pLDLR–LDLR expression plasmids led to a significant reduction in plasmid LDL cholesterol, which was further enhanced by co-injection with miRNA expression vectors targeted to mouse Hmgcr. Our data suggest that targeted knockdown of HMGCR may enhance gene therapy outcomes for FH.

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Acknowledgements

This work was supported by the British Heart Foundation grants PG/08/028/24799 and PG/09/029/27185. We thank Dr Zoe Holloway and Dr Natalia Sacilotto for their kind gifts of Hep3b and Hepa 1-6 cells. We thank Dr Rebecca Fairclough for her assistance with imaging and Ms Emma Carter for her technical assistance with plasma cholesterol measurements.

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Authors and Affiliations

  1. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK

    O Hibbitt, S Agkatsev, C Owen, M Cioroch & R Wade-Martins

  2. Department of Clinical Pharmacology, University of Oxford, Oxford, UK

    L Seymour

  3. Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UK

    K Channon

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  1. O Hibbitt
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  2. S Agkatsev
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Correspondence to R Wade-Martins.

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Hibbitt, O., Agkatsev, S., Owen, C. et al. RNAi-mediated knockdown of HMG CoA reductase enhances gene expression from physiologically regulated low-density lipoprotein receptor therapeutic vectors in vivo. Gene Ther 19, 463–467 (2012). https://doi.org/10.1038/gt.2011.103

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