HPMA polymer-based site-specific delivery of oligonucleotides to hepatic stellate cells.
The objective was to determine whether bioconjugation of type I collagen specific triplex forming oligonucleotide (TFO) to N-(2-hydroxypropyl) methacrylamide (HPMA) containing tetrapeptide Gly-Phe-Leu-Gly (GFLG) and mannose 6-phosphate (M6P) can provide their targeted delivery to hepatic stellate cells (HSCs). Following bioconjugation, M6P-GFLG-HPMA-GFLG-32P-TFO was characterized by PAGE, HPLC, and GPC, and then its biodistribution was determined. TFO was dissociated from the conjugate when incubated with papain and formed triplex with the target DNA duplex. Type 1 collagen gene expression was significantly inhibited when HSC-T6 cells were transfected with this conjugate. Following tail vein injection into rats, M6P-GFLG-HPMA-GFLG-(32)P-TFO was rapidly cleared from the circulation and accumulated mainly in the liver. The plasma concentration versus time profile was biphasic, with 12.37 min as t(1/2) of distribution and 2886.48 min as t(1/2) of elimination. A large proportion of the injected M6P-GFLG-HPMA-GFLG-32P-TFO was taken up by the HSCs of both normal and fibrotic rats, which were isolated by liver perfusion at 30 min post-injection. Preinjection of M6P-GFLG-HPMA-GFLG-ONP into fibrotic rats decreased the liver uptake of the conjugates from 60% to 13%, suggesting M6P/TGFII receptor-mediated endocytosis of the conjugates by HSCs. Almost 80% of the total liver uptake in fibrotic rats was contributed by HSCs. In conclusion, conjugation with M6P-HPMA-GFLG significantly increased TFO delivery to the HSCs and could be potentially used for treating liver fibrosis.
Yang
Ningning
N
Department of Pharmaceutical Sciences, University of Tennessee, Health Science Center, Memphis, Tennessee 38103, USA.
Ye
Zhaoyang
Z
Li
Feng
F
Mahato
Ram I
RI
D000178:Acrylamides / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry* / Q000378:metabolism*; D005355:Fibrosis / Q000378:metabolism / Q000473:pathology; D055166:Hepatic Stellate Cells / Q000378:metabolism*; D007267:Injections; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008360:Mannosephosphates / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D014018:Tissue Distribution
D000178:Acrylamides; D024042:Collagen Type I; D004337:Drug Carriers; D008360:Mannosephosphates; D009841:Oligonucleotides; D009842:Oligopeptides; C051802:triplex DNA; C027693:mannose-6-phosphate; D004247:DNA; C032976:N-(2-hydroxypropyl)methacrylamide