Double oxygen-sensing vector system for robust hypoxia/ischemia-regulated gene induction in cardiac muscle in vitro and in vivo.

Abstract

High-fidelity genetically encoded bio-sensors that respond to changes in cellular environmental milieu in disease offer great potential in a range of patho-physiological settings. Here a unique hypoxia-regulated vector-based system with double oxygen-sensing transcriptional elements was developed for rapid and robust hypoxia-regulated gene expression in the heart. Hypoxia-responsive cis elements were used in tandem with a single proline-modified oxygen-dependent degradation (ODD) domain of hypoxia-inducible factor-1alpha to form a double oxygen-sensing vector system (DOSVS). In adult cardiac myocytes in vitro, the DOSVS demonstrated a low background expression not different from baseline control in normoxia, and with 100% efficiency, robust, 1,000-fold induction upon hypoxia. In the heart in vivo, hypoxic and ischemic challenges elicited rapid 700-fold induction in living animals, exceeding that obtained by a high-fidelity constitutive cytomegalovirus (CMV) viral promoter. DOSVS also showed high temporal resolution in the heart in response to cyclical bouts of hypoxia in vivo. We propose that DOSVS will be valuable for a range of applications, including bio-sensing and therapeutic gene expression in the heart and other organ systems that are confronted by chronic or episodic hypoxic/ischemic stresses in vivo.

• Authors: Ekaterina V Fomicheva, Immanuel I Turner, Terri G Edwards, Janet Hoff, Eric Arden, Louis G D'Alecy, Joseph M Metzger
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 16 Issue 9 Pg. 1594-601 (Sep 2008) ISSN: 1525-0024 [Electronic] United States
• PMID: 18578010 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• enhanced green fluorescent protein
• Green Fluorescent Proteins
• Luciferases
• Oxygen

Topics

• Animals
• Cells, Cultured
• Cytomegalovirus (genetics)
• Gene Expression Regulation
• Genetic Vectors
• Green Fluorescent Proteins (genetics)
• Humans
• Hypoxia (genetics, therapy)
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics, therapeutic use)
• In Vitro Techniques
• Ischemia (genetics, therapy)
• Luciferases (metabolism)
• Myocardium (metabolism, pathology)
• Myocytes, Cardiac (metabolism, pathology)
• Oxygen (metabolism)
• Promoter Regions, Genetic (genetics)
• Rats
• Rats, Sprague-Dawley
• Response Elements (genetics)
• Transcriptional Activation
• Transduction, Genetic