Highly Selective Targeting of Hepatic Stellate Cells for Liver Fibrosis Treatment Using a d-Enantiomeric Peptide Ligand of Fn14 Identified by Mirror-Image mRNA Display

Abstract

Although liver fibrosis is a major public health issue, there is still no effective drug therapy in the clinic. Fibroblast growth factor-inducible 14 (Fn14), a membrane receptor highly specifically expressed in activated hepatic stellate cells (HSCs), is the key driver of liver fibrosis, and thus, it has a great potential as a novel target for the development of effective treatment. Here, we identified a d-enantiomeric peptide ligand of Fn14 through mirror-image mRNA display. This included the chemical synthesis of a d-enantiomer of the target protein (extracellular domain of Fn14), identification of an l-peptide ligand of d-Fn14 using a constructed mRNA peptide library, and identification of a d-enantiomer of the l-peptide, which is a ligand of the natural Fn14 for reasons of symmetry. The obtained d-peptide ligand showed strong binding to Fn14 while maintaining high proteolytic resistance. As a targeting moiety, this d-peptide successfully mediated high selectivity of activated HSCs for liposomal vehicles compared to that of other major cell types in the liver and significantly enhanced the accumulation of liposomes in the liver fibrosis region of a carbon tetrachloride-induced mouse model. Moreover, in combination with curcumin as an encapsulated load, a liposomal formulation conjugated with this d-peptide showed powerful inhibition of the proliferation of activated HSCs and reduced the liver fibrosis to a significant extent in vivo. This Fn14-targeting strategy may represent a promising approach to targeted drug delivery for liver fibrosis treatment. Meanwhile, the mirror-image mRNA display can provide a new arsenal for the development of d-peptide-based therapeutics against a variety of human diseases.

pPB Peptide-Mediated siRNA-Loaded Stable Nucleic Acid Lipid Nanoparticles on Targeting Therapy of Hepatic Fibrosis

“…Currently, some new therapeutic approaches have been developed to reverse hepatic fibrosis. , For example, it is reported that the formation of hepatic fibrosis could be suppressed by inhibiting the expression of cell-associated collagen. Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone in the endoplasmic reticulum .…”

Selective deletion of MyD88 signaling in α-SMA positive cells ameliorates experimental intestinal fibrosis via post-transcriptional regulation

“…While α-SMA cell selective strategies are not yet available, mesenchymal cell targeted approaches have been tested in experimental liver fibrosis using the platelet-derived growth factor beta receptor or fibroblast growth factor-inducible 14 (Fn14). 58 , 59 With the development of new endpoints for clinical trials in fibrostenosing CD the implementation of this strategy may become reality. 60 …”

In-cell penetration selection—mass spectrometry produces noncanonical peptides for antisense delivery

“…29–31 Again, biological display techniques are restricted to the use of mostly natural amino acids, limiting the resulting library diversity and proteolytic stability 32,33 , and even those mirror image techniques that allow D-peptide discovery still have difficulty incorporating non-canonical residues. 34,35 Screening of a synthetic one-bead one-compound (OBOC) library eases the incorporation of non-canonical and D-residues. Our group has recently demonstrated that in vivo affinity selection-mass spectrometry (AS-MS) could identify an erythrocyte-targeting D-peptide.…”