Receptor-mediated hepatic uptake of M6P-BSA-conjugated triplex-forming oligonucleotides in rats.
Excessive production of extracellular matrix, predominantly type I collagen, results in liver fibrosis. Earlier we synthesized mannose 6-phosphate-bovine serum albumin (M6P-BSA) and conjugated to the type I collagen specific triplex-forming oligonucleotide (TFO) for its enhanced delivery to hepatic stellate cells (HSCs), which is the principal liver fibrogenic cell. In this report, we demonstrate a time-dependent cellular uptake of M6P-BSA-33P-TFO by HSC-T6 cells. Both cellular uptake and nuclear deposition of M6P-BSA-33P-TFO were significantly higher than those of 33P-TFO, leading to enhanced inhibition of type I collagen transcription. Following systemic administration into rats, hepatic accumulation of M6P-BSA-33P-TFO increased from 55% to 68% with the number of M6P per BSA from 14 to 27. Unlike 33P-TFO, there was no significant decrease in the hepatic uptake of (M6P)20-BSA-33P-TFO in fibrotic rats. Prior administration of excess M6P-BSA decreased the hepatic uptake of (M6P)20-BSA-33P-TFO from 66% to 40% in normal rats, and from 60% to 15% in fibrotic rats, suggesting M6P/insulin-like growth factor II (M6P/IGF II) receptor-mediated endocytosis of M6P-BSA-33P-TFO by HSCs. Almost 82% of the total liver uptake in fibrotic rats was contributed by HSCs. In conclusion, by conjugation with M6P-BSA, the TFO could be potentially used for the treatment of liver fibrosis.
Ye
Zhaoyang
Z
Department of Pharmaceutical, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
Cheng
Kun
K
Guntaka
Ramareddy V
RV
Mahato
Ram I
RI
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects; D024042:Collagen Type I / Q000235:genetics; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism*; D008360:Mannosephosphates / Q000737:chemistry* / Q000378:metabolism; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology*; D051381:Rats; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D014158:Transcription, Genetic / Q000235:genetics
D002352:Carrier Proteins; D024042:Collagen Type I; D008360:Mannosephosphates; D009841:Oligonucleotides; D012710:Serum Albumin, Bovine; C027693:mannose-6-phosphate