Development of a Gene Delivery System of Oligonucleotides for Fibroses by Targeting Cell-Surface Vimentin-Expressing Cells with N-Acetylglucosamine-Bearing Polymer-Conjugated Polyethyleneimine

Abstract

<jats:p>Targeting myofibroblasts and activated stellate cells in lesion sites of fibrotic tissues is an important approach to treat fibroses. Herein, we focused on targeting the cytoskeletal proteins vimentin, which are reportedly highly expressed on the surface of these cells and have N-acetylglucosamine (GlcNAc)-binding activity. A GlcNAc-bearing polymer synthesized via radical polymerization with a reversible addition-fragmentation chain transfer reagent has been previously found to interact with cell-surface vimentin-expressing cells. We designed a GlcNAc-bearing polymer-conjugated polyethyleneimine (PEI), as the gene carrier to target cell-surface vimentin-expressing cells and specifically deliver nuclear factor-κB decoy oligonucleotides (ODNs) and heat shock protein 47 (HSP47)-small interfering RNA (siRNA) to normal human dermal fibroblasts (NHDFs) that express cell-surface vimentin. The results showed that the expression of tumor necrosis factor-α in lipopolysaccharide-stimulated NHDFs and HSP47 in transforming growth factor-β1-stimulated NHDFs was suppressed by cellular uptake of the GlcNAc-bearing polymer-conjugated PEI/nuclear factor (NF)-κB decoy ODNs and HSP47-siRNA complexes through cell-surface vimentin, respectively. These findings suggest that the effective and specific delivery of ODNs and siRNA for cell-surface vimentin-expressing cells such as myofibroblasts and activated stellate cells can be achieved using GlcNAc-bearing polymer-conjugated PEI. This therapeutic approach could prove advantageous to prevent the promotion of various fibroses.</jats:p>

Comparison of circulating tumor cell (CTC) detection rates with epithelial cell adhesion molecule (EpCAM) and cell surface vimentin (CSV) antibodies in different solid tumors: a retrospective study

“…The reason for the false-positive results with EpCAM method might be due to the potential skin epithelial cell contamination from venipuncture. CSV cannot only be seen in EMT type of cancer cells, but also in apoptotic neutrophils, some virus-infected cells and in myofibroblasts and activated stellate cells ( Moisan & Girard, 2006 ; Du et al, 2014 ; Song & Ise, 2020 ). CSV expression is shown to have correlations with metastatic phenotype in aggressive cancer and sarcoma ( Satelli & Li, 2011 ; Satelli et al, 2014 ; Satelli et al, 2015 ).…”

“…As a putative anti-viral drug target, CSV takes part in viral multiplication by facilitating the internalization of virions ( Du et al, 2014 ). Therefore, false-positive results with CSV method might be observed in people with inflammatory diseases, virus infection or fibrosis lesion ( Moisan & Girard, 2006 ; Du et al, 2014 ; Song & Ise, 2020 ). Vimentin is normally expressed in mesenchymal cells, such as white blood cells (WBC), as a component of cytoskeleton ( Satelli & Li, 2011 ).…”

“…EpCAM is not expressed in every epithelial tumor ( ) and may be lost in cells undergoing EMT ( Chaw et al, 2012 ). CSV is expressed in EMT cells, apoptotic neutrophils, virus-infected cells and myofibroblasts ( Satelli & Li, 2011 ; Moisan & Girard, 2006 ; Du et al, 2014 ; Song & Ise, 2020 ). The use of CSV to detect CTCs in the blood may generate some false-positive results.…”

Evaluation of Cell Surface Vimentin Positive Circulating Tumor Cells as a Diagnostic Biomarker for Lung Cancer

“…CSV has been identified to participate in cell adhesion, migration and cellular signaling (21). Expression of CSV is not only seen in cells undergoing EMT, but also in cells infected with certain virus, and in activated lymphocytes, myofibroblasts and stellate cells (40)(41)(42). As inflammation and fibrosis are usually common observed in patients with BLD, the false positives might come from the activated lymphocytes and myofibroblasts.…”

Peer Review #1 of "Comparison of circulating tumor cell (CTC) detection rates with epithelial cell adhesion molecule (EpCAM) and cell surface vimentin (CSV) antibodies in different solid tumors: a retrospective study (v0.1)"

“…The reason for the false-positive results with EpCAM method might be due to the potential skin epithelial cell contamination from venipuncture. CSV cannot only be seen in EMT type of cancer cells, but also in apoptotic neutrophils, some virusinfected cells and in myofibroblasts and activated stellate cells [33][34][35]. CSV expression is shown to have correlations with metastatic phenotype in aggressive cancer and sarcoma [19][20][21].…”