Heme oxygenase-1 fused to a TAT peptide transduces and protects pancreatic beta-cells.
Transplantation of islets is becoming an established method for treating type 1 diabetes. However, viability of islets is greatly affected by necrosis/apoptosis induced by oxidative stress and other insults during isolation and subsequent in vitro culture. Expression of cytoprotective proteins, such as heme oxygenase-1 (HO-1), reduces the deleterious effects of oxidative stress in transplantable islets. We have generated a fusion protein composed of HO-1 and TAT protein transduction domain (TAT/PTD), an 11-aa cell penetrating peptide from the human immunodeficiency virus TAT protein. Transduction of TAT/PTD-HO-1 to insulin-producing cells protects against TNF-alpha-mediated cytotoxicity. TAT/PTD-HO-1 transduction to islets does not impair islet physiology, as assessed by reversion of chemically induced diabetes in immunodeficient mice. Finally, we report that transduction of HO-1 fusion protein into islets improves islet viability in culture. This approach might have a positive impact on the availability of islets for transplantation.
Ribeiro
Melina M
MM
Diabetes Research Institute, University of Miami School of Medicine, 1450 NW 10th Avenue (R-134), Miami, FL 33136, USA.
Klein
Dagmar
D
Pileggi
Antonello
A
Molano
R Damaris
RD
Fraker
Christopher
C
Ricordi
Camillo
C
Inverardi
Luca
L
Pastori
Ricardo L
RL
D000595:Amino Acid Sequence; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D019610:Cytoprotection; D015696:Gene Products, tat / Q000737:chemistry*; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics*; D051547:Heme Oxygenase-1; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D010455:Peptides / Q000737:chemistry; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D011917:Rats, Inbred Lew; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D015696:Gene Products, tat; D007328:Insulin; D010455:Peptides; D011993:Recombinant Fusion Proteins; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1