Introduction 1 2 3 4 Pdx1 Neurod1 Btc 5 8 7 5 8 9 10 9 11 13 14 15 16 17 18 Materials and methods Vector production 18 10 19 20 9 g 5 21 Transduction of hepatocytes in vitro 2+ 2+ 2+ 2+ 3 4 4 2 4 3 22 5 7 Transduction of hepatocytes in vivo 8 Microscopic analyses 2 23 Functional studies 23 24 25 PCR analyses T Qualitative western blot analyses Statistical analyses t Results Efficient transduction of hepatocytes p 3 1 1 5 Fig. 1 a b c d e  1 1 Transduction with HMD/INS-FUR reversed STZ-diabetes  2 2 Fig. 2 a b black circles white squares black triangles n  HMD/INS-FUR transduction stimulated expression of pancreatic hormones 3 3 3 3 3 Fig. 3 a b c d e f 2 2 2 g h i  3 3 HMD/INS-FUR transduction induced hepatic insulin production n 4 Fig. 4 Arrows  HMD/INS-FUR transduced animals displayed regulated insulin secretion 5 5 5 Fig. 5 a b c White squares black diamonds black triangles n  HMD/INS-FUR transduction stimulated expression of beta cell transcription factors 6 6 6 Fig. 6 a b Pdx1, Neurod1, Neurog3, Nkx2-2, Nkx6-1, Pax4, Pax6 Sst Gcg Ppy Slc2a2 Gck Ins1 Ins2 Pc2 P48 Actb c  Pdx1 Neurod1 6 Neurog3 Nkx2-2 3 Ins1 Ins2 Pc2 Pcsk2 Ptf1a P48 6 Pdx1 Neurod1 Neurog3 p Pdx1 NeuroD1 Neurog3 Discussion This is the first report of permanent amelioration of STZ-induced hyperglycaemia using LV delivery of INS-FUR. Transduced animals processed, stored and secreted insulin in a glucose-regulated manner. Remission of diabetes, due to the secretion of insulin from residual beta cells, was discounted as insulin-positive pancreatic beta cells were rarely observed and levels of rat C-peptide were negligible. This is also the first report showing that transduction of INS-FUR alone can induce expression of several beta cell transcription factors resulting in hepatocyte differentiation along a pancreatic lineage. 23 26 5 8 27 28 Pdx1 Neurod1 29 30 32 23 PDX1 5 7 33 34 Pdx1 35 36 Pdx1 37 38 39 40 5 7 41 18 42 43 44 45 19 46 47 48 49 50 9 Pdx1 28