HIF-regulated HO-1 gene transfer improves the post-ischemic limb recovery and diminishes TLR-triggered immune responses - Effects modified by concomitant VEGF overexpression

Agnieszka Jazwa; Mateusz Stoszko; Mateusz Tomczyk; Karolina Bukowska-Strakova; Chantal Pichon; Alicja Jozkowicz; Jozef Dulak

doi:10.1016/j.vph.2015.02.011

HIF-regulated HO-1 gene transfer improves the post-ischemic limb recovery and diminishes TLR-triggered immune responses - Effects modified by concomitant VEGF overexpression

Vascul Pharmacol. 2015 Aug:71:127-38. doi: 10.1016/j.vph.2015.02.011. Epub 2015 Apr 10.

Authors

Agnieszka Jazwa¹, Mateusz Stoszko², Mateusz Tomczyk², Karolina Bukowska-Strakova³, Chantal Pichon⁴, Alicja Jozkowicz², Jozef Dulak⁵

Affiliations

¹ Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland. Electronic address: [email protected].
² Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
³ Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; Department of Clinical Immunology and Transplantology, Polish-American Institute of Pediatrics, Medical College, Jagiellonian University, Krakow, Poland.
⁴ Centre de Biophysique Moleculaire, CNRS, Orleans, France; University of Orleans, Orleans, France.
⁵ Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland; Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.

PMID: 25869523
DOI: 10.1016/j.vph.2015.02.011

Abstract

Heme oxygenase-1 (HO-1) mitigates cellular injury by antioxidant, anti-apoptotic, anti-inflammatory and proangiogenic effects. Vascular endothelial growth factor (VEGF) is a critical regulator of blood vessel growth. Their coordinated action was analyzed in a model of femoral artery ligation (FAL) in mice lacking HO-1 gene (HO-1 KO). Gastrocnemius skeletal muscles of HO-1 KO mice were preemptively injected with plasmids containing hypoxia-response element (HRE) driving the expression of only HO-1 (pHRE-HO1) or both HO-1 and VEGF (pHRE-HO1-VEGF). At day 14th the pHRE-HO1 vector increased an impaired post-ischemic blood flow recovery in HO-1 KO mice to the level observed in wild-type (WT) mice subjected to FAL and pHRE-HO1-VEGF restored it already at day 7. The pHRE-HO1 gene therapy diminished, when compared to control pHRE-empty-treated HO-1 KO mice, the expression of toll-like receptors (TLR4 and TLR9) and inflammatory cytokines (IL-1β, IL-6 and TNFα) at day 3, whereas opposite effects were observed following concomitant HO-1 and VEGF gene transfer. Moreover, HO-1 diminished ischemia-induced expression of MyoD involved in satellite cell differentiation in HO-1 KO mice. Our results confirm the therapeutic potential of HO-1 and VEGF against critical limb ischemia although, their concomitant delivery may have contradictory actions on the resolution of inflammation.

Keywords: Gene therapy; HO-1; HRE; Inflammation; Regeneration; TLR; VEGF.

Publication types

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

MeSH terms

Animals
Female
Gene Expression Regulation
Gene Transfer Techniques
Heme Oxygenase-1 / administration & dosage
Heme Oxygenase-1 / genetics*
Hindlimb / blood supply
Hypoxia-Inducible Factor 1, alpha Subunit / administration & dosage
Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
Immunity, Cellular / drug effects
Immunity, Cellular / physiology*
Ischemia / genetics*
Ischemia / immunology
Ischemia / therapy
Male
Mice
Mice, Inbred C57BL
Mice, Knockout
Recovery of Function / physiology
Toll-Like Receptors / biosynthesis*
Toll-Like Receptors / immunology
Vascular Endothelial Growth Factor A / biosynthesis*

Substances

Hif1a protein, mouse
Hypoxia-Inducible Factor 1, alpha Subunit
Toll-Like Receptors
Vascular Endothelial Growth Factor A
vascular endothelial growth factor A, mouse
HMOX1 protein, human
Heme Oxygenase-1