Establishment of gene therapy targeted for renal mesangial and proximal tubular cells

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(1) Development of single-chane monoclonoal antibody against mesangial cellsTo develop single-chain monoclonal antibody against mesangial cells, Balb/c mouse were immunized with rat and human mesangial cells. From these immunized mouse spleen, CDNA library of phage-display type monoclonal antibody was developed. Currently, second-screening of this library is being performed, although specific antibody has not been obtained, yet.(2) Gene transfer of truncated IκBα to renal proximal cells in renal impairment ratsAs renal interstitial injury model, albumin-loaded rats were used. Gene transfer of truncated IκBα, which inhibits nuclear factor κB (NFκB), key molecule for inflammation, was performed with recombinant adenovirus via renal artery. Adenovirus transferred reporter gene into proximal cells, efficiently, and transferred truncated IκBα abolished tubulo-interstitial changes by albumin loading. From these results, it was suggested that inhibition of NFκB prevented renal damage in this model, we also examined a possible drug for the treatment of renal diseases. We found that, tranilast, which is known to inhibit NFκB in several cell lines, could inhibit NFκB activation in mesangial cells, suggesting that this drug may be useful for clinical settings.To express transferred gene more efficiently, we analyzed promoter activity of vitamin D 1α-hydroxylase, which is predominantly expressed in proximal tubules. This study showed that region of 105 bp upstream of the initiation codon has promoter activity for expression in proximal tubular cell line, LLC-PK1. Furthermore, TCF-1 binding motif in this region is shown to play important roles in promoter activity.