Cationic liposome-mediated gene transfer during acute pancreatitis: tissue specificity, duration, and effects of acute inflammation.
Production of inflammatory cytokines in the pancreas, lung, and liver is believed to play a major role in the development of severe pancreatitis. This tissue-specific production could lend itself to directed anti-cytokine gene therapy if an appropriate delivery system could be developed. This study was undertaken to examine a novel approach for the delivery of protein-based therapies to the tissues involved during acute pancreatitis. Healthy mice received an intraperitoneal injection of cationic liposomes and a DNA plasmid containing the chloramphenicol acetyltransferase (CAT) reporter gene. Animals were killed at 12 hours and 1, 2, 3, 7, and 14 days with serum, pancreas, lung, and liver harvested. Acute pancreatitis was induced (cerulein, 50 micrograms/kg/hr intraperitoneally x4) in additional mice before or after CAT transfection. The presence of pancreatitis was established in all animals by histologic scoring of pancreata and by serum amylase and lipase levels. CAT transfection efficiency was determined by quantitative CAT enzyme activity within tissue homogenates. Animals that received the liposome were successfully transfected with the CAT gene into the pancreas, lungs, and liver. Maximal transfection in each tissue occurred at 12 hours with decreasing CAT activity over the ensuing 14 days. No healthy animals receiving the CAT gene developed elevations in amylase, lipase, or any histologic parameter of pancreatitis. Transfection efficiency in the pancreas was markedly increased by preexisting or delayed induction of pancreatitis, whereas transfection of the lung and liver was increased to a lesser extent. Gene transfection into the pancreas, liver, and lungs is possible using a cationic liposome delivery system that does not induce pancreatitis or pancreatic inflammation. Pancreatic expression of the gene product is equal to or greater than that of the organs of the reticuloendothelial system and continues at very high efficiency rates during acute pancreatitis.
Denham
W
W
Department of Surgery, University of South Florida, Tampa, FL, USA.
Yang
J
J
MacKay
S
S
Tannahill
C
C
Carter
G
G
Abouhamze
A
A
Moldawer
L L
LL
Norman
J
J
D000208:Acute Disease; D000681:Amylases / Q000097:blood; D000818:Animals; D002412:Cations; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000097:blood / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D004195:Disease Models, Animal; D005500:Follow-Up Studies; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D008049:Lipase / Q000097:blood; D008081:Liposomes; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D051379:Mice; D010179:Pancreas / Q000201:enzymology / Q000473:pathology; D010195:Pancreatitis / Q000097:blood / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D010957:Plasmids; D013045:Species Specificity; D013997:Time Factors; D014162:Transfection
D002412:Cations; D008081:Liposomes; D015500:Chloramphenicol O-Acetyltransferase; D008049:Lipase; D000681:Amylases