Regulated expression of a foreign gene targeted to the ischaemic myocardium.
OBJECTIVE
Regulated expression of transferred foreign genes may be an important feature of gene therapy. Because coronary artery disease often involves intermittent myocardial ischaemia followed by periods of normal cardiac function it will probably be necessary to regulate the expression of putative therapeutic/cardioprotective genes directly in response to ischaemia-associated signals. The objectives of the current study were to develop a combination of gene regulatory components that can be used to target a product to the myocardium and limit the expression of the gene to periods of ischaemic activity.


METHODS
Expression plasmids were constructed containing muscle-specific promoters and hypoxia-responsive enhancer elements linked to a reporter gene. The regulation of these constructs by hypoxia or experimental ischaemia was measured following transient expression in cultured cells or after direct injection of DNA into the rabbit myocardium.


RESULTS
A single set of hypoxia response elements placed immediately upstream of the minimal muscle-specific alpha-myosin heavy chain promoter conferred potent positive regulation of this promoter by hypoxia in vitro and by ischaemia in vivo. Induction by ischaemia persisted for at least 4 h and returned to the baseline level within 8 h.


CONCLUSIONS
Hypoxia responsive regulatory elements, in combination with weak tissue-restricted promoters incorporated into an appropriate vector system may allow controlled expression of a therapeutic gene in ischaemic myocardium.
Prentice
H
H
Pharmaceutical Discovery Division, SRI International, Menlo Park, CA, USA.
Bishopric
N H
NH
Hicks
M N
MN
Discher
D J
DJ
Wu
X
X
Wylie
A A
AA
Webster
K A
KA
D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005809:Genes, Regulator; D015316:Genetic Therapy / Q000379:methods*; D000860:Hypoxia / Q000235:genetics; D008297:Male; D017202:Myocardial Ischemia / Q000235:genetics / Q000628:therapy*; D009206:Myocardium; D018995:Myosin Heavy Chains / Q000235:genetics; D010084:Oxidation-Reduction; D011817:Rabbits; D051381:Rats
D018995:Myosin Heavy Chains