Spatial and temporal control of transgene expression through ultrasound-mediated induction of the heat shock protein 70B promoter in vivo.
Adenoviral vectors have been constructed that express the transgenes luciferase (Adeno-HSP-Luc) or Fas ligand (Adeno-HSP-FasL) under the control of the heat shock protein 70B (hsp70B) promoter. Cultures infected with Adeno-HSP-Luc transiently expressed high levels of luciferase after heat shock. When cultures infected with Adeno-HSP-FasL were maintained at 37 degrees C, no transgene expression was observed, but when cultures were exposed to heat stress, transgene expression resulted in apoptotic cell death. In vivo, transgene expression was induced by ultrasound-mediated heating of adenovirus-infected tissue. In mice or rats injected with the Adeno-HSP-Luc construct, high levels of localized expression of luciferase activity were observed in regions subjected to ultrasound-mediated irradiation. Adeno-HSP-FasL was administered systemically to mice via the tail vein to evaluate safety. Animals receiving Adeno-HSP-FasL in the absence of ultrasound treatment did not display liver toxicity, whereas animals receiving ultrasound treatment to induce the expression of Fas ligand from the hsp70B promoter had significant increases in serum levels of liver enzymes. These data demonstrate that combining the inducible hsp70B promoter with ultrasound induction allows safe local expression of cytotoxic genes with possible therapeutic utility.
Smith
Roy C
RC
Molecular Cardiology/CVI, Whitaker Cardiovascular Institute, Boston University School of Medicine, 75 Albany Street, W611, Boston, MA 02118, USA. rcsmith@bu.edu
Machluf
Marcelle
M
Bromley
Peter
P
Atala
Anthony
A
Walsh
Kenneth
K
D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D053222:Fas Ligand Protein; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008811:Mice, Inbred DBA; D011401:Promoter Regions, Genetic; D019076:Transgenes
C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D018840:HSP70 Heat-Shock Proteins; C491129:HSPA7 protein, human; D008562:Membrane Glycoproteins; D008156:Luciferases