Protection of islets in culture by delivery of oxygen binding neuroglobin via protein transduction.
Islet transplantation has become an accepted method to treat type 1 diabetes. To succeed and achieve normal levels of glucose in transplant recipients, the quality of the transplanted islets is of the utmost importance. Lack of oxygen during organ procurement, islet isolation, and subsequent culture triggers apoptosis or necrosis and loss of islet function, causing the yield and quality to diminish. A promising candidate for cytoprotection against oxygen deprivation is neuroglobin (Ngb). Ngb is a recently described member of globin family and is expressed in neurons, retina, and pancreatic islets. To overexpress this protein in the islets and study its ability to protect them, we utilized protein transduction. Protein transduction is achieved by fusing Ngb to the TAT/PTD transduction domain, a peptide originated from the HIV transcriptional transactivator protein. Our study proved that TAT-Ngb is an efficient fusion protein capable of protecting the human islets in culture from loss of cell mass and function, thus increasing the quality of transplantable islets. If the islets could be cultured for a longer period of time without suffering harmful effects, it would be possible to precondition the recipient and there would be more time to assess their quality and function before transplantation.
Mendoza
V
V
Diabetes Research Institute, University of Miami School of Medicine, Miami, Florida 33136, USA.
Klein
D
D
Ichii
H
H
Ribeiro
M M
MM
Ricordi
C
C
Hankeln
T
T
Burmester
T
T
Pastori
R L
RL
D001331:Automation; D015687:Cell Hypoxia; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D005434:Flow Cytometry; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism; D005914:Globins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology; D016381:Islets of Langerhans Transplantation / Q000502:physiology; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D000078787:Neuroglobin; D010101:Oxygen Consumption; D021381:Protein Transport
D015696:Gene Products, tat; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D005914:Globins