Epigenetic upregulation of endogenous VEGF-A reduces myocardial infarct size in mice

Mikko P Turunen; Tiia Husso; Haja Musthafa; Svetlana Laidinen; Galina Dragneva; Nihay Laham-Karam; Sanna Honkanen; Anne Paakinaho; Johanna P Laakkonen; Erhe Gao; Maija Vihinen-Ranta; Timo Liimatainen; Seppo Ylä-Herttuala

doi:10.1371/journal.pone.0089979

Epigenetic upregulation of endogenous VEGF-A reduces myocardial infarct size in mice

PLoS One. 2014 Feb 26;9(2):e89979. doi: 10.1371/journal.pone.0089979. eCollection 2014.

Affiliations

¹ Department of Biotechnology and Molecular Medicine, A.I.Virtanen Institute, University of Eastern Finland, Kuopio, Finland.
² The Center for Translational Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania, United States of America.
³ Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylä, Finland.
⁴ Department of Biotechnology and Molecular Medicine, A.I.Virtanen Institute, University of Eastern Finland, Kuopio, Finland ; Research Unit and Gene Therapy Unit, Kuopio University Hospital, Kuopio, Finland.

Abstract

"Epigenetherapy" alters epigenetic status of the targeted chromatin and modifies expression of the endogenous therapeutic gene. In this study we used lentiviral in vivo delivery of small hairpin RNA (shRNA) into hearts in a murine infarction model. shRNA complementary to the promoter of vascular endothelial growth factor (VEGF-A) was able to upregulate endogenous VEGF-A expression. Histological and multiphoton microscope analysis confirmed the therapeutic effect in the transduced hearts. Magnetic resonance imaging (MRI) showed in vivo that the infarct size was significantly reduced in the treatment group 14 days after the epigenetherapy. Importantly, we show that promoter-targeted shRNA upregulates all isoforms of endogenous VEGF-A and that an intact hairpin structure is required for the shRNA activity. In conclusion, regulation of gene expression at the promoter level is a promising new treatment strategy for myocardial infarction and also potentially useful for the upregulation of other endogenous genes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
Cyclic AMP Response Element-Binding Protein / metabolism
DNA Methylation
Epigenesis, Genetic*
Gene Silencing
Inverted Repeat Sequences / genetics
Male
Membrane Proteins / metabolism
Mice
Mice, Inbred C57BL
Myocardial Infarction / genetics*
Myocardial Infarction / metabolism
Myocardial Infarction / pathology*
Phosphoproteins / metabolism
Promoter Regions, Genetic / genetics
Protein Isoforms / genetics
RNA, Small Interfering / genetics
Transcription, Genetic / genetics
Transcriptional Activation
Up-Regulation / genetics*
Vascular Endothelial Growth Factor A / genetics*

Substances

Cyclic AMP Response Element-Binding Protein
Membrane Proteins
Pag1 protein, mouse
Phosphoproteins
Protein Isoforms
RNA, Small Interfering
Vascular Endothelial Growth Factor A

Grants and funding

This study was supported by grants from Finnish Academy, ERC, Kuopio University Hospital, Sigrid Juselius Foundation, Finnish Foundation for Cardiovascular Research, Jenny and Antti Wihuri Foundation, Eye and Tissue bank Foundation, Orion-Farmos Research Foundation and Instrumentarium Science Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.