Transplantation of Gunn rats with autologous fibroblasts expressing bilirubin UDP-glucuronosyltransferase: correction of genetic deficiency and tumor formation.
The end product of the breakdown of the heme group of hemoglobin and other heme-containing proteins is bilirubin. Bilirubin is hydrophobic and cannot be excreted as such. Therefore, mammals have a liver enzyme bilirubin UDP-glucuronosyltransferase (B-UGT), which conjugates bilirubin with glucuronic acid, thereby making the molecule much more water soluble. Bilirubin glucuronides are secreted into bile. Patients with Crigler-Najjar (CN) disease have a deficiency in bilirubin UDP-glucuronosyltransferase and accumulate high serum levels of bilirubin. An animal model for CN disease is the Gunn rat. The obvious target for gene therapy for CN disease is the liver, but because liver cells do only divide infrequently, they are difficult to transduce. To investigate whether cells that are easily transduced can be used to develop gene therapy for CN disease, we have transduced Gunn rat fibroblasts with B-UGT, using a recombinant retrovirus. Gunn rat fibroblasts expressing B-UGT were able to glucuronidate bilirubin present in cell culture media. In this study, we describe the intraperitoneal transplantation of Gunn rats with Gunn rat fibroblasts expressing B-UGT. Transplantation of the fibroblasts corrected the genetic deficiency of the Gunn rats, serum bilirubin concentrations of the transplanted Gunn rats were reduced to normal, and bilirubin glucuronides appeared in bile. However, due to the prolonged period of cell culture, the transplanted fibroblasts were transformed, and the experimental animals developed tumors after transplantation.
Seppen
J
J
Department of Gastrointestinal and Liver Diseases, Academic Medical Centre, Amsterdam, The Netherlands.
Tada
K
K
Ottenhoff
R
R
Sengupta
K
K
Chowdhury
N R
NR
Chowdhury
J R
JR
Bosma
P J
PJ
Oude Elferink
R P
RP
D000818:Animals; D001646:Bile / Q000737:chemistry / Q000378:metabolism; D001663:Bilirubin / Q000097:blood; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003414:Crigler-Najjar Syndrome / Q000235:genetics / Q000628:therapy; D004195:Disease Models, Animal; D005347:Fibroblasts; D015870:Gene Expression / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D005966:Glucuronidase / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism*; D007150:Immunohistochemistry; D009374:Neoplasms, Experimental / Q000378:metabolism; D051381:Rats; D011911:Rats, Gunn; D017208:Rats, Wistar; D012190:Retroviridae / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics; D014182:Transplantation, Autologous
D014453:Glucuronosyltransferase; D005966:Glucuronidase; D001663:Bilirubin