ADENOVIRUS-MEDIATED GENE-TRANSFER INTO HUMAN HEPATOCYTES - ANALYSIS OF THE BIOCHEMICAL FUNCTIONALITY OF TRANSDUCED CELLS

The use of replication-defective adenoviruses to deliver transgenes in to hepatocytes seems to be a promising approach to human liver gene th erapy. However, the effects that the adenovirus-mediated expression of a foreign gene could have on the expression of other hepatic characte ristic genes have not yet been properly examined. We have investigated this problem by using human hepatocytes infected with a recombinant E -1 defective adenovirus that that carried a modified lacZ gene. The an alysis of the biochemical functionality of transduced cells showed tha t the use of adenovirus: (1) was a very efficient way to introduce a f oreign gene into human hepatocytes (80% transduced cells after 1 h con tact, at an MOI of 15; approximately 100% transduced cells at an MOI o f 20); (2) allowed the expression of the transgene to levels that enab led cells effectively to use lactose as an energy source; (3) does not affect urea synthesis, plasma protein synthesis and xenobiotic biotra nsformation activities (1A2, 2A1, 2B6, 3A3/5). Glycolysis was moderate ly increased (approximately 20%); while gluconeogenesis decreased (app roximately 20%) in transduced hepatocyte; moreover, (4) the expression of inducible genes (acute-phase plasma proteins, CYPs) was not impair ed in transduced human hepatocytes upon stimulation with IL-6 or methy lcholantrene. The results of this research support the idea that effic ient expression of transgenes can be achieved in human hepatocytes by means of adenoviral transduction, without altering these characteristi c hepatic biochemical functions.