GFAP promoter-driven RNA interference on TGF-β1 to treat liver fibrosis

Ningning Yang, Ram I. Mahato

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Purpose: The objective was to determine the role of promoters and miRNA backbone in shRNA-based hepatic stellate cell (HSC)-specific transforming growth factor (TGF)-β1 gene silencing. This is expected to avoid the side effect of non-specific TGF-β1 gene silencing. Methods: Two most potent shRNAs targeting 769 and 1033 start sites of rat TGF-β1 mRNA were cloned into pSilencer 1.0 vector for enhanced TGF-β1 gene silencing. We then constructed HSC-specific pri-miRNA mimic and pri-miRNA cluster mimic expression plasmids in which shRNA expression was driven by a glial fibrillary acidic protein (GFAP) promoter to achieve HSC-specific TGF-β1 gene silencing to avoid nonspecific inhibition of TGF-β1 expression in other cells and organs. Results: These TGF-β1 pri-miRNA-producing plasmids showed the inhibition of proliferation and induced apoptosis of activated HSC-T6 cells. TGF-β1 pri-miRNA cluster mimic plasmids decreased TGF-β1 and collagen gene expression at both mRNA and protein levels. Conclusions: GFAP promoter driven TGF-β1 pri-miRNA producing plasmids have the potential to be used for site-specific gene therapeutics to treat liver fibrosis.

Original languageEnglish (US)
Pages (from-to)752-761
Number of pages10
JournalPharmaceutical Research
Volume28
Issue number4
DOIs
StatePublished - Apr 1 2011

Fingerprint

Glial Fibrillary Acidic Protein
Transforming Growth Factors
RNA Interference
Liver Cirrhosis
Liver
RNA
MicroRNAs
Hepatic Stellate Cells
Gene Silencing
Genes
Plasmids
Small Interfering RNA
Messenger RNA
Gene expression
Rats
Collagen
Apoptosis
Gene Expression

Keywords

  • GFAP promoter
  • TGF-β1
  • liver fibrosis
  • pri-miRNA mimics
  • shRNA

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry
  • Pharmacology (medical)

Cite this

GFAP promoter-driven RNA interference on TGF-β1 to treat liver fibrosis. / Yang, Ningning; Mahato, Ram I.

In: Pharmaceutical Research, Vol. 28, No. 4, 01.04.2011, p. 752-761.

Research output: Contribution to journalArticle

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