Treatment with human metalloproteinase-8 gene delivery ameliorates experimental rat liver cirrhosis.
OBJECTIVE
An extrahepatic human neutrophil collagenase complementary DNA (matrix metalloprotease-8) cloned in an adenovirus vector was used as a therapeutic agent in cirrhosis.


METHODS
A high titer of clinical-grade AdMMP8 was obtained.


RESULTS
HeLa cells transduced with AdMMP8 expressed recombinant matrix metalloprotease-8 messenger RNA and matrix metalloprotease-8 protein. Matrix metalloprotease-8 in culture sups showed enzymatic activity against native collagen type I, which was inhibited by ethylenediaminetetraacetic acid, 1,10-phenanthroline, and tissue inhibitor of metalloprotease-1. In vivo transduction showed matrix metalloprotease-8 activity, and studies to establish the efficacy of this characterized vector were performed in CCl(4) and bile duct-ligated cirrhotic rats. Transduction with 3 x 10(11) viral particles per kilogram resulted in hepatic detection of both messenger RNA and protein matrix metalloprotease-8. A consistent response in fibrosis reversal was observed in CCl(4) rats. Liver fibrosis in bile duct-ligated cirrhotic animals was decreased in 45%, along with diminished hydroxyproline content, after AdMMP8 treatment. The expression of matrix metalloprotease-2 and matrix metalloprotease-3 was up-regulated in AdMMP8 rats. Free tissue inhibitor of metalloprotease-1, as an indirect measurement of active uncomplexed matrix metalloproteases, was also increased in the AdMMP8 groups. Transforming growth factor-beta messenger RNA was diminished, and matrix metalloprotease-9 and hepatocyte growth factor increased. Treatment in both models correlated with improvements in ascites, functional hepatic tests, and gastric varices, indicating diminished intrahepatic blood pressure in animals injected with AdMMP8.


CONCLUSIONS
Therefore, therapy with the matrix metalloprotease-8 gene is promising for use in a clinical setting.
Siller-López
Fernando
F
Institute for Molecular Biology in Medicine and Gene Therapy, Centro Universitario de Ciencias de la Salud, University of Guadalajara, Mexico.
Sandoval
Ana
A
Salgado
Silvia
S
Salazar
Adriana
A
Bueno
Miriam
M
Garcia
Jesus
J
Vera
Jose
J
Gálvez
Javier
J
Hernández
Iván
I
Ramos
Martha
M
Aguilar-Cordova
Estuardo
E
Armendariz-Borunda
Juan
J
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002251:Carbon Tetrachloride; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007084:Iliac Vein; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000628:therapy*; D008297:Male; D020784:Matrix Metalloproteinase 8 / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar
D003470:Culture Media; D012333:RNA, Messenger; D003094:Collagen; D002251:Carbon Tetrachloride; D020784:Matrix Metalloproteinase 8