Vitamin A-coupled liposomes carrying TLR4-silencing shRNA induce apoptosis of pancreatic stellate cells and resolution of pancreatic fibrosis

Abstract

Chronic pancreatitis leads to irreversible damage in pancreatic endocrine and exocrine functions. However, there is no clinically available antifibrotic drug. Pancreatic stellate cells (PSCs) can be activated by Toll-like receptor 4 (TLR4) responses to its ligands and they contribute to the formation of pancreatic fibrosis. Silencing the expression of TLR4 in PSCs by RNAi may be a novel therapeutic strategy for the treatment of pancreatic fibrosis. In addition, PSCs have a remarkable capacity for vitamin A uptake most likely through cellular retinol binding protein (CRBP). In our study, to ensure the efficient delivery of RNAi therapeutic agents to PSCs, VitA-coupled liposomes (VA-lips) were used as drug carriers to deliver plasmids expressing TLR4-specific short hairpin RNA (shRNA) to treat pancreatic fibrosis. Our study demonstrated that silencing the expression of TLR4 could induce mitochondrial apoptosis in aPSCs and might be an effective therapeutic strategy for the treatment of pancreatic fibrosis.

Key messages

  • VA-lip-shRNA-TLR4 recovers pancreatic tissue damage.

  • VA-lip-shRNA-TLR4 resolution of pancreatic fibrosis.

  • VA-lip-shRNA-TLR4 accelerates ECM degradation and inhibits ECM synthesis.

  • Silencing TLR4 induces aPSCs mitochondrial apoptosis.

  • Silencing TLR4 inhibits the activation of NF-κB.

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Funding

This work was supported by the National Natural Science Foundation of China (Grant No. 81400665).

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Correspondence to Ping Cheng.

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The authors declare no conflict of interest related to this study.

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Supplementary Fig. 1

Evaluation of the side effects of VA-lip-shRNA-TLR4. a Representative images of H&E staining for liver (top panel) and lung (bottom panel) (× 200). b Alanine aminotransferase and c aspartate aminotransferase levels in the sera of mice. Values are the mean ± SD for each group (n = 10 per group). (JPEG 2933 kb)

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Zhang, Y., Yue, D., Cheng, L. et al. Vitamin A-coupled liposomes carrying TLR4-silencing shRNA induce apoptosis of pancreatic stellate cells and resolution of pancreatic fibrosis. J Mol Med 96, 445–458 (2018). https://doi.org/10.1007/s00109-018-1629-6

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Keywords

  • Pancreatic fibrosis
  • Pancreatic stellate cells
  • Toll-like receptor 4
  • Gene therapy
  • RNAi
  • Liposome