Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells

Zhaoyang Ye, Kun Cheng, Ramareddy Guntaka, Ram I. Mahato

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Liver fibrosis is characterized by abnormal accumulation of extracellular matrix (ECM), namely, fibrillar collagens in the hepatic stellate cells (HSCs). Earlier, we developed an antigene approach, using a type α1(I) collagen gene promoter specific triplex-forming oligonucleotide (TFO) to inhibit collagen gene expression. In this paper, to enhance overall delivery of TFOs to the liver and more specifically to HSCs, we synthesized man nose 6-phosphate-bovine serum albumin (M6P-BSA) by phosphorylating p-nitrophenyl-α-D- mannopyranoside, reducing its nitro group, and reacting it with thiophosgene to produce p-isothiocyanatophenyl-6-phospho-α-D-mannopyranoside (itcM6P) for conjugation with BSA. 33P-TFO was conjugated with M6P-BSA via a disulfide bond, and the stability of the (M6P)20-BSA-TFO conjugate was determined. Following tail vein injection into rats, (M6P) 20-BSA-33P-TFO rapidly cleared from the circulation and accumulated mainly in the liver. Almost 66% of the injected (M6P) 20-BSA33P-TFO accumulated in the liver at 30 min postinjection, which was significantly higher than that deposited after injection of 33P-TFO. A large proportion of the injected (M6P) 20-BSA-33P-TFO was taken up by the HSCs as evidenced by determination of radioactivity in the digested liver cells upon liver perfusion and separation on a Nycodenz gradient. Therefore, this TFO conjugate may be used for the treatment of liver fibrosis.

Original languageEnglish (US)
Pages (from-to)4466-4476
Number of pages11
JournalBiochemistry
Volume44
Issue number11
DOIs
StatePublished - Mar 22 2005

Fingerprint

Hepatic Stellate Cells
Oligonucleotides
Liver
Mannose
Bovine Serum Albumin
Nose
Liver Cirrhosis
Collagen
Phosphates
Fibrillar Collagens
Iohexol
Injections
Radioactivity
Collagen Type I
Gene expression
Disulfides
Extracellular Matrix
Tail
Rats
Veins

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells. / Ye, Zhaoyang; Cheng, Kun; Guntaka, Ramareddy; Mahato, Ram I.

In: Biochemistry, Vol. 44, No. 11, 22.03.2005, p. 4466-4476.

Research output: Contribution to journalArticle

Ye, Zhaoyang ; Cheng, Kun ; Guntaka, Ramareddy ; Mahato, Ram I. / Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells. In: Biochemistry. 2005 ; Vol. 44, No. 11. pp. 4466-4476.
@article{6fcbd3da5d4b405bba22431e1bcf51f3,
title = "Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells",
abstract = "Liver fibrosis is characterized by abnormal accumulation of extracellular matrix (ECM), namely, fibrillar collagens in the hepatic stellate cells (HSCs). Earlier, we developed an antigene approach, using a type α1(I) collagen gene promoter specific triplex-forming oligonucleotide (TFO) to inhibit collagen gene expression. In this paper, to enhance overall delivery of TFOs to the liver and more specifically to HSCs, we synthesized man nose 6-phosphate-bovine serum albumin (M6P-BSA) by phosphorylating p-nitrophenyl-α-D- mannopyranoside, reducing its nitro group, and reacting it with thiophosgene to produce p-isothiocyanatophenyl-6-phospho-α-D-mannopyranoside (itcM6P) for conjugation with BSA. 33P-TFO was conjugated with M6P-BSA via a disulfide bond, and the stability of the (M6P)20-BSA-TFO conjugate was determined. Following tail vein injection into rats, (M6P) 20-BSA-33P-TFO rapidly cleared from the circulation and accumulated mainly in the liver. Almost 66{\%} of the injected (M6P) 20-BSA33P-TFO accumulated in the liver at 30 min postinjection, which was significantly higher than that deposited after injection of 33P-TFO. A large proportion of the injected (M6P) 20-BSA-33P-TFO was taken up by the HSCs as evidenced by determination of radioactivity in the digested liver cells upon liver perfusion and separation on a Nycodenz gradient. Therefore, this TFO conjugate may be used for the treatment of liver fibrosis.",
author = "Zhaoyang Ye and Kun Cheng and Ramareddy Guntaka and Mahato, {Ram I.}",
year = "2005",
month = "3",
day = "22",
doi = "10.1021/bi047529j",
language = "English (US)",
volume = "44",
pages = "4466--4476",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "11",

}

TY - JOUR

T1 - Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells

AU - Ye, Zhaoyang

AU - Cheng, Kun

AU - Guntaka, Ramareddy

AU - Mahato, Ram I.

PY - 2005/3/22

Y1 - 2005/3/22

N2 - Liver fibrosis is characterized by abnormal accumulation of extracellular matrix (ECM), namely, fibrillar collagens in the hepatic stellate cells (HSCs). Earlier, we developed an antigene approach, using a type α1(I) collagen gene promoter specific triplex-forming oligonucleotide (TFO) to inhibit collagen gene expression. In this paper, to enhance overall delivery of TFOs to the liver and more specifically to HSCs, we synthesized man nose 6-phosphate-bovine serum albumin (M6P-BSA) by phosphorylating p-nitrophenyl-α-D- mannopyranoside, reducing its nitro group, and reacting it with thiophosgene to produce p-isothiocyanatophenyl-6-phospho-α-D-mannopyranoside (itcM6P) for conjugation with BSA. 33P-TFO was conjugated with M6P-BSA via a disulfide bond, and the stability of the (M6P)20-BSA-TFO conjugate was determined. Following tail vein injection into rats, (M6P) 20-BSA-33P-TFO rapidly cleared from the circulation and accumulated mainly in the liver. Almost 66% of the injected (M6P) 20-BSA33P-TFO accumulated in the liver at 30 min postinjection, which was significantly higher than that deposited after injection of 33P-TFO. A large proportion of the injected (M6P) 20-BSA-33P-TFO was taken up by the HSCs as evidenced by determination of radioactivity in the digested liver cells upon liver perfusion and separation on a Nycodenz gradient. Therefore, this TFO conjugate may be used for the treatment of liver fibrosis.

AB - Liver fibrosis is characterized by abnormal accumulation of extracellular matrix (ECM), namely, fibrillar collagens in the hepatic stellate cells (HSCs). Earlier, we developed an antigene approach, using a type α1(I) collagen gene promoter specific triplex-forming oligonucleotide (TFO) to inhibit collagen gene expression. In this paper, to enhance overall delivery of TFOs to the liver and more specifically to HSCs, we synthesized man nose 6-phosphate-bovine serum albumin (M6P-BSA) by phosphorylating p-nitrophenyl-α-D- mannopyranoside, reducing its nitro group, and reacting it with thiophosgene to produce p-isothiocyanatophenyl-6-phospho-α-D-mannopyranoside (itcM6P) for conjugation with BSA. 33P-TFO was conjugated with M6P-BSA via a disulfide bond, and the stability of the (M6P)20-BSA-TFO conjugate was determined. Following tail vein injection into rats, (M6P) 20-BSA-33P-TFO rapidly cleared from the circulation and accumulated mainly in the liver. Almost 66% of the injected (M6P) 20-BSA33P-TFO accumulated in the liver at 30 min postinjection, which was significantly higher than that deposited after injection of 33P-TFO. A large proportion of the injected (M6P) 20-BSA-33P-TFO was taken up by the HSCs as evidenced by determination of radioactivity in the digested liver cells upon liver perfusion and separation on a Nycodenz gradient. Therefore, this TFO conjugate may be used for the treatment of liver fibrosis.

UR - http://www.scopus.com/inward/record.url?scp=15544382666&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=15544382666&partnerID=8YFLogxK

U2 - 10.1021/bi047529j

DO - 10.1021/bi047529j

M3 - Article

VL - 44

SP - 4466

EP - 4476

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 11

ER -