PCBP2 siRNA Reverses the Alcohol-induced Pro-fibrogenic Effects in Hepatic Stellate Cells

Shukla, Ravi; Qin, Bin; Wan, Yu‐Jui Yvonne; Cheng, Kun

doi:10.1007/s11095-011-0475-9

Cited by 17 publications

(42 citation statements)

References 45 publications

(66 reference statements)

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“…Cells were passaged at 80%–90% confluence. For alcohol stimulation experiments, HSCs were incubated with alcohol in a closed chamber as described previously [9, 16–18]. …”

Section: Methodsmentioning

confidence: 99%

“…Cells were transfected using Lipofectamine-iMAX as described previously [9, 19, 20]. The cells were seeded in 24-well plates at a density of 10,000–30,000 cells/well at 12 h before the transfection.…”

Section: Methodsmentioning

confidence: 99%

“…The excessive collagen accumulation in activated HSC is mainly due to the enhanced stability of type I collagen mRNA [8]. In our previous study [9], HSC-T6 cells exposed to alcohol for 48 h demonstrated enhanced αCP2 expression leading to the prolonged stability of type I collagen mRNA.…”

Section: Introductionmentioning

confidence: 99%

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Silencing of α-complex protein-2 reverses alcohol- and cytokine-induced fibrogenesis in hepatic stellate cells

et al. 2017

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Background and aim α-complex protein-2 (αCP2) encoded by the poly (rC) binding protein 2(PCBP2) gene is responsible for the accumulation of type I collagen in fibrotic livers. In this study, we silenced the PCBP2 gene using a small interfering RNA (siRNA) to reverse alcohol-and cytokine-induced profibrogenic effects on hepatic stellate cells (HSCs). Methods Primary rat HSCs and the HSC-T6 cell line were used as fibrogenic models to mimic the initiation and perpetuation stages of fibrogenesis, respectively. We previously found that a PCBP2 siRNA, which efficiently silences expression of αCP2, reduces the stability of type I collagen mRNA. We investigated the effects of the PCBP2 siRNA on cell proliferation and migration. Expression of type I collagen in HSCs was analyzed by quantitative real-time PCR and western blotting. In addition, we evaluated the effects of the PCBP2 siRNA on apoptosis and the cell cycle. Results PCBP2 siRNA reversed multiple alcohol- and cytokine-induced profibrogenic effects on primary rat HSCs and HSC-T6 cells. The PCBP2 siRNA also reversed alcohol- and cytokine-induced accumulation of type I collagen as well as cell proliferation and migration. Moreover, the combination of LY2109761, a transforming growth factor-β1 inhibitor, and the PCBP2 siRNA exerted a synergistic inhibitive effect on the accumulation of type I collagen in HSCs. Conclusions Silencing of PCBP2 using siRNA could be a potential therapeutic strategy for alcoholic liver fibrosis.

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“…Cells were passaged at 80%–90% confluence. For alcohol stimulation experiments, HSCs were incubated with alcohol in a closed chamber as described previously [9, 16–18]. …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Silencing of α-complex protein-2 reverses alcohol- and cytokine-induced fibrogenesis in hepatic stellate cells

et al. 2017

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“…[1, 2] Various therapeutic agents including small molecular antifibrotic molecules, oligonucleotides and siRNA have been studied for treating liver fibrosis. [3–5] For example, we recently developed an siRNA targeting the poly(rC) binding protein 2 (PCBP2) gene, which is overexpressed and responsible for the stabilization of the collagen α1(I) mRNA in liver fibrogenesis. [5] However, the siRNA has to be specifically delivered to its target cells to exert its therapeutic effect with minimum toxicity in other tissues.…”

Section: Introductionmentioning

confidence: 99%

“…[3–5] For example, we recently developed an siRNA targeting the poly(rC) binding protein 2 (PCBP2) gene, which is overexpressed and responsible for the stabilization of the collagen α1(I) mRNA in liver fibrogenesis. [5] However, the siRNA has to be specifically delivered to its target cells to exert its therapeutic effect with minimum toxicity in other tissues. Activation of hepatic stellate cells (HSCs) is the crucial step of fibrogensis because HSCs are the main producers responsible for the excessive production of ECM and profibrogenic cytokines in fibrotic liver.…”

Section: Introductionmentioning

confidence: 99%

Discovery of Peptide Ligands for Hepatic Stellate Cells Using Phage Display

et al. 2015

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Regardless of its cause, liver fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM) in the liver. Hepatic stellate cells (HSCs) are the main producers responsible for the excessive production of ECM and profibrogenic cytokines in fibrotic liver. Therefore, development of HSC-specific delivery systems is essential for the success of antifibrotic agents. The objective of this study is to identify peptide ligands targeting the insulin like growth factor 2 receptor (IGF2R), which is overexpressed on HSCs. We expect to use the peptide ligands for the future development of HSC-targeted drug delivery system. Protein- and whole cell-based phage display biopannings were conducted to identify phage/peptide candidates. Phage ELISA, cellular uptake and cell viability assay were employed to evaluate the binding affinity and specificity of these peptide ligands to recombinant human IGF2R and HSCs. IGF2R siRNA was used to silence the IGF2R protein expression in human hepatic stellate cells (LX-2) to confirm the specificity of the identified peptide ligands. Among the identified peptide candidates, the peptide-431 shows the highest binding affinity and specificity to recombinant human IGF2R protein and HSCs. The equilibrium dissociation constant (Kd) of the peptide-431 is 6.19 μM for LX-2 cells and 12.35 μM for rat hepatic stellate cells HSC-T6. Cellular uptake of the peptide-431 in LX-2 cells is significantly reduced after silencing IGF2R with siRNA. The peptide-431 also enhances the uptake of a proapoptotic peptide (KLA peptide) in LX-2 and HSC-T6 cells, indicating that the peptide-431 can be used as a targeting ligand to deliver antifibrotic agents into not only rat but also human HSCs. Dimerization of the peptide-431 further increase its binding affinity to LX-2 cells by approximately nine-fold.

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Targeted Delivery of an siRNA/PNA Hybrid Nanocomplex Reverses Carbon Tetrachloride‐Induced Liver Fibrosis

Jain

Barve

Zhao

et al. 2019

Advanced Therapeutics

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Liver fibrosis is a wound healing process marked by excessive accumulation of extracellular matrix in the liver. A poly(rC)-binding protein 2 (PCBP2) siRNA that reverses fibrogenesis in activated hepatic stellate cells (HSCs) has been recently discovered. However, targeted delivery of siRNAs to HSCs still remains a daunting challenge. Herein, a new strategy is developed to fabricate a multicomponent nanocomplex using siRNA/peptide nucleic acid (PNA) hybrid instead of chemically conjugated siRNA, thus increasing the scalability and feasibility of the siRNA nanocomplex for animal studies. The nanocomplex is modified with an insulin growth factor 2 receptor -specific peptide, which specifically binds to activated HSCs. The siRNA nanocomplex demonstrates a controllable size, high serum stability, and high cellular uptake in activated HSCs in vitro and in vivo. Anti-fibrotic activity of the siRNA nanocomplex is evaluated in rats with carbon tetrachloride-induced liver fibrosis. Treatment with the PCBP2 siRNA nanocomplex significantly inhibits the mRNA expressions of PCBP2 and type I collagen in fibrotic liver. The histology study reveals that the siRNA nanocomplex efficiently reduces the protein level of type I collagen and reverses liver fibrosis. The data suggests that the nanocomplex efficiently delivers the siRNA to fibrotic liver and produces a potent anti-fibrotic effect.

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PCBP2 siRNA Reverses the Alcohol-induced Pro-fibrogenic Effects in Hepatic Stellate Cells

Cited by 17 publications

References 45 publications

Silencing of α-complex protein-2 reverses alcohol- and cytokine-induced fibrogenesis in hepatic stellate cells

Silencing of α-complex protein-2 reverses alcohol- and cytokine-induced fibrogenesis in hepatic stellate cells

Discovery of Peptide Ligands for Hepatic Stellate Cells Using Phage Display

Targeted Delivery of an siRNA/PNA Hybrid Nanocomplex Reverses Carbon Tetrachloride‐Induced Liver Fibrosis

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