HPMA polymer-based site-specific delivery of oligonucleotides to hepatic stellate cells

Ningning Yang, Zhaoyang Ye, Feng Li, Ram I. Mahato

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

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- 32P-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 t1/2 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.

Original languageEnglish (US)
Pages (from-to)213-221
Number of pages9
JournalBioconjugate Chemistry
Volume20
Issue number2
DOIs
StatePublished - Feb 1 2009

Fingerprint

Hepatic Stellate Cells
Oligonucleotides
Polymers
Phosphates
Liver
Rats
Collagen Type I
Gly-Phe-Leu-Gly
Collagen
IGF Type 2 Receptor
Papain
Injections
Endocytosis
Gene expression
Liver Cirrhosis
N-(2-hydroxypropyl)methacrylamide
mannose-6-phosphate
Tail
Veins
Perfusion

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

HPMA polymer-based site-specific delivery of oligonucleotides to hepatic stellate cells. / Yang, Ningning; Ye, Zhaoyang; Li, Feng; Mahato, Ram I.

In: Bioconjugate Chemistry, Vol. 20, No. 2, 01.02.2009, p. 213-221.

Research output: Contribution to journalArticle

Yang, Ningning ; Ye, Zhaoyang ; Li, Feng ; Mahato, Ram I. / HPMA polymer-based site-specific delivery of oligonucleotides to hepatic stellate cells. In: Bioconjugate Chemistry. 2009 ; Vol. 20, No. 2. pp. 213-221.
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abstract = "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- 32P-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 t1/2 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.",
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