Volume 272, Number 9,
Issue of February 28, 1997
pp. 5800-5804
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
The Pancreatitis-associated Protein I Promoter Allows Targeting
to the Pancreas of a Foreign Gene, Whose Expression Is Up-regulated
during Pancreatic Inflammation
(Received for publication, September 10, 1996, and in revised form, December 5, 1996)
Nelson J.
Dusetti
,
Sophie
Vasseur
,
Emilia M.
Ortiz
,
Horacio
Romeo
,
Jean-Charles
Dagorn
,
Oscar
Burrone
**
and
Juan L.
Iovanna
From 
U.315 INSERM, 46 boulevard de la Gaye, F 13009 Marseille, France and the ** International Centre for Genetic
Engineering and Biotechnology, Area Science Park, Padriacino
99, 34012 Trieste, Italy
The pancreatitis-associated protein I
(PAP I) is a pancreatic secretory protein expressed in pancreas during
acute pancreatitis but not in the healthy pancreas. The promoter of the
PAP I gene thus represents a potential candidate to drive expression of
therapeutic molecules to the diseased pancreas. In this work, we have
constructed recombinant adenoviruses harboring the chloramphenicol
acetyltransferase (CAT) gene driven by several fragments of the PAP I
promoter and have characterized their properties in vitro
and in vivo. In vitro studies showed that the
transduction of the pancreatic cell line AR-42J with these adenoviruses
led to low levels of CAT activity in basal conditions. After
stimulation with a combination of interleukin-6 and dexamethasone
or after induction of oxidative stress, CAT activity was strongly
induced, a characteristic of the endogenous PAP I gene. Stimulation was
maximal when constructs comprised 1253 base pairs of the PAP I
promoter, upstream from initiation of transcription, and decreased with
shorter fragments of 317, 180, 118 or 61 base pairs. The recombinant
adenovirus containing the CAT gene under the control of the PAP I
promoter fragment (
1253/+10) was also tested in vivo.
Following administration by intravenous injection into mice, CAT
activity was measured in several tissues 96 h later. In healthy
animals, low but significant CAT activity was detected in pancreas,
compared with near background values observed in the other tissues.
When experimental acute pancreatitis was induced, CAT expression was
strongly enhanced only in pancreas. In control experiments with
adenoviruses in which the CAT gene was driven by the cytomegalovirus
promoter, higher levels of expression were observed in all tissues.
Expression was not modified after induction of acute pancreatitis. In
conclusion, this study shows that (i) a recombinant adenovirus
containing a fragment of the PAP I promoter allows specific targeting
of a reporter gene to the mouse pancreas and (ii) expression of the reporter gene in pancreas is induced during acute pancreatitis. Adenovirus-mediated gene therapy of acute pancreatitis is therefore conceivable.
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