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

Ekaterina V. Fomicheva, Immanuel I. Turner, Terri G. Edwards, Janet Hoff, Eric Arden, Louis G. D'Alecy, Joseph M. Metzger

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

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-1α 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.

Original languageEnglish (US)
Pages (from-to)1594-1601
Number of pages8
JournalMolecular Therapy
Volume16
Issue number9
DOIs
StatePublished - 2008

Bibliographical note

Funding Information:
We thank David Pinsky, Diane Bouis, and Hui Liao for assistance and access with their hypoxia station and Steven Whitesall for help with hypoxic-conditioning experiments. We also thank Eric Devaney and Jackie Cale for helpful comments on an earlier version of this manuscript and Jennifer Davis for assistance with the figures. This work was supported by an National Institutes of Health grant to J.M.M. and a postdoctoral fellowship grant from The American Heart Association to Ekaterina Fomicheva (0620089Z).

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