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Construction of minicircle DNA vectors capable of correcting familial hypercholesterolemia phenotype in a LDLR-deficient mouse model

Gene Therapy volume 23, pages 657–663 (2016)Cite this article

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Abstract

Familial hypercholesterolemia (FH) caused by defect in low-density lipoprotein receptor (LDLR) is a life-threatening disease with poor response to conventional treatments. Earlier gene therapy studies have generated promising results, but further development is hampered because the cells harboring the viral vectors were eliminated by host immune system soon after delivery, whereas the nonviral vectors were too bulky to be delivered to target cells. To overcome these problems, we constructed multiple minicircle (MC) DNA vectors to express the therapeutic LDLR. MC is an optimized nonviral vector that is capable of expressing high level of transgene product persistently. We found that among the seven MCs tested, the best is MC5 with multiple advanced features. First, the LDLr gene was placed under the control of sterol regulatory element (SRE) using LDLr gene promoter or apoprotein E (ApoE) promoter, allowing the transcription of the LDLr gene to be regulated by serum low-density lipoprotein (LDL) cholesterol as its functional gene counterpart. Second, a hepatic control region (HCR) was placed upstream of the promoter that serves as a controller to ensure liver-specific expression. Third, the modified Kozak sequence was placed in front of the LDLr gene start codon to enhance its translation efficiency. MC5 was 5.23 kb in size, and was capable of tight physiological control in intracellular LDL cholesterol level even when challenged with high dose of sterols in vitro. Importantly, it was able to correct the phenotype of LDLR-deficient mice C57BL/6 LDLR-/- for more than 105 days without detectable toxicity. Therefore, this MC has the clinical application potential for treating FH.

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Acknowledgements

This work is funded by the Shenzhen Government (Grant Numbers SFG 2012566 and SKC 2012237).

Author contributions

Z-YC perceived the concept and worked with XHH and RJ to design the experiments and write the manuscript. XHH, RJ, TYW, XYG, YSC and C-YH conducted the experiments.

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Author notes

  1. R Jiao: This author contributed equally to this work.

Authors and Affiliations

  1. The Laboratory for Gene and Cell Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China

    X Hou, R Jiao, X Guo, T Wang, P Chen, D Wang, Y Chen, C-Y He & Z-Y Chen

  2. Department of Food Science and Engineering, Jinan University, Guangzhou, China

    R Jiao

  3. Department of Infectious Diseases, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China

    X Guo

Authors
  1. X Hou
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  5. P Chen
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  7. Y Chen
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Correspondence to Z-Y Chen.

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The authors declare no conflict of interest.

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Hou, X., Jiao, R., Guo, X. et al. Construction of minicircle DNA vectors capable of correcting familial hypercholesterolemia phenotype in a LDLR-deficient mouse model. Gene Ther 23, 657–663 (2016). https://doi.org/10.1038/gt.2016.37

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