Targeted Delivery of an siRNA/PNA Hybrid Nanocomplex Reverses Carbon Tetrachloride-Induced Liver Fibrosis
Akshay Jain1, Ashutosh Barve1, Zhen Zhao1
1Division of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, Kansas City, MO 64108, USA.
Abstract:
Liver fibrosis is a wound healing process with excessive accumulation of extracellular matrix in the liver. We recently discovered a PCBP2 siRNA that reverses fibrogenesis in activated hepatic stellate cells (HSCs), which are the key players in liver fibrogenesis. However, targeted delivery of siRNAs to HSCs still remains a challenge. Herein, we developed a new strategy to fabricate a multicomponent nanocomplex using siRNA/PNA hybrid instead of chemically conjugated siRNA, thus increasing the scalability and feasibility of the siRNA nanocomplex for animal studies. We modified the nanocomplex with an insulin growth factor 2 receptor (IGF2R)-specific peptide, which specifically binds to activated HSCs. The siRNA nanocomplex shows a controllable size and high serum stability. The nanocomplex also demonstrates high cellular uptake in activated HSCs in vitro and in vivo. Anti-fibrotic activity of the siRNA nanocomplex was 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. Histology study revealed that the siRNA nanocomplex efficiently reduces the protein level of type I collagen and reverses liver fibrosis. Our data suggest that the nanocomplex efficiently delivers the siRNA to fibrotic liver and produces a potent anti-fibrotic effect.
Insights
This study developed a novel nanocomplex for targeted delivery of PCBP2 siRNA to liver cells, effectively reversing liver fibrosis in rats. The innovative approach enhances therapeutic potential for liver fibrogenesis treatment.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Hepatology
Background:
- Liver fibrosis involves excessive extracellular matrix accumulation driven by activated hepatic stellate cells (HSCs).
- Targeted delivery of small interfering RNA (siRNA) to HSCs is crucial for treating liver fibrosis but remains challenging.
- PCBP2 siRNA has shown potential to reverse fibrogenesis in activated HSCs.
Purpose of the Study:
- To develop a novel, scalable, and feasible siRNA nanocomplex for targeted delivery to activated HSCs.
- To evaluate the anti-fibrotic efficacy of a PCBP2 siRNA nanocomplex in a rat model of liver fibrosis.
Main Methods:
- Fabrication of a multicomponent nanocomplex using an siRNA/PNA hybrid, modified with an insulin growth factor 2 receptor (IGF2R)-specific peptide for HSC targeting.
- In vitro and in vivo assessment of nanocomplex cellular uptake and serum stability.
- Evaluation of anti-fibrotic activity in rats with carbon tetrachloride-induced liver fibrosis, measuring PCBP2 and type I collagen expression and histological changes.
Main Results:
- The developed siRNA nanocomplex exhibited controllable size, high serum stability, and efficient cellular uptake in activated HSCs both in vitro and in vivo.
- Treatment with the PCBP2 siRNA nanocomplex significantly reduced PCBP2 and type I collagen mRNA expression in fibrotic livers.
- Histological analysis confirmed a significant reduction in type I collagen protein levels and reversal of liver fibrosis.
Conclusions:
- The novel nanocomplex effectively delivers PCBP2 siRNA to fibrotic liver tissue.
- The PCBP2 siRNA nanocomplex demonstrates potent anti-fibrotic effects, offering a promising therapeutic strategy for liver fibrosis.
