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Cirrhotic rat livers with extensive fibrosis can be safely transduced with clinical-grade adenoviral vectors. Evidence of cirrhosis reversion

Gene Therapy volume 9, pages 127–134 (2002)Cite this article

Abstract

Adenoviral vectors efficiently target normal liver cells; however, a clear-cut description of the safety boundaries for using adenovectors in hepatic cirrhosis has not been settled. With this in mind, we used a first-generation, replication-deficient adenoviral vector carrying the E. coli lacZ gene (Ad5βGal) to monitor therapeutic range, biodistribution, toxicity and transduction efficiency in Wistar rats made cirrhotic by two different experimental approaches resembling alcoholic cirrhosis and biliary cirrhosis in humans. Further, we show proof of concept on fibrosis reversion by a ‘therapeutic’ Ad-vector (AdMMP8) carrying a gene coding for a collagen-degrading enzyme. Dose–response experiments with Ad5βGal ranging from 1 × 108–3 × 1012 viral particles (vp) per rat (250 g), demonstrated that adenovirus-mediated gene transfer via iliac vein at 3 × 1011 vp/rat, resulted in an approximately 40% transduction in livers of rats made cirrhotic by chronic intoxication with carbon tetrachloride, compared with approximately 80% in control non-cirrhotic livers. In rats made cirrhotic by bile-duct obstruction only, 10% efficiency of transduction was observed. Biodistribution analyses showed that vector expression was detected primarily in liver and at a low level in spleen and kidney. Although there was an important increase in liver enzymes between the first 48 h after adenovirus injection in cirrhotic animals compared to non-transduced cirrhotic rats, this hepatic damage was resolved after 72–96 h. Then, the cDNA for neutrophil collagenase, also known as Matrix Metalloproteinase 8 (MMP8), was cloned in an Ad-vector and delivered to cirrhotic rat livers being able to reverse fibrosis in 44%. This study demonstrates the potential use of adenoviral vectors in safe transient gene therapy strategies for human liver cirrhosis.

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Acknowledgements

This work was supported in part by a grant from CONACyT No. 28832-M to Juan Armendariz-Borunda. The authors are indebted to Dr Guillermo Grijalva and Jose M Vera for their invaluable assistance.

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Authors and Affiliations

  1. Institute of Molecular Biology in Medicine and Gene Therapy, CUCS, University of Guadalajara, Guadalajara, Mexico

    J Garcia-Bañuelos, F Siller-Lopez, A Miranda & J Armendariz-Borunda

  2. Advantagene, Inc., San Diego, CA, USA

    L K Aguilar

  3. Harvard Gene Therapy Initiative, Boston, MA, USA

    E Aguilar-Cordova

Authors
  1. J Garcia-Bañuelos
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  2. F Siller-Lopez
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  3. A Miranda
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  6. J Armendariz-Borunda
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Garcia-Bañuelos, J., Siller-Lopez, F., Miranda, A. et al. Cirrhotic rat livers with extensive fibrosis can be safely transduced with clinical-grade adenoviral vectors. Evidence of cirrhosis reversion. Gene Ther 9, 127–134 (2002). https://doi.org/10.1038/sj.gt.3301647

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