Ameliorating liver fibrosis in an animal model using the secretome released from miR-122-transfected adipose-derived stem cells

Abstract

BACKGROUNDRecently, the exclusive use of mesenchymal stem cell (MSC)-secreted molecules, called secretome, rather than cells, has been evaluated for overcoming the limitations of cell-based therapy, while maintaining its advantages. However, the use of naïve secretome may not fully satisfy the specificity of each disease. Therefore, it appears to be more advantageous to use the functionally reinforced secretome through a series of processes involving physico-chemical adjustments or genetic manipulation rather than to the use naïve secretome.AIMTo determine the therapeutic potential of the secretome released from miR-122-transfected adipose-derived stromal cells (ASCs).METHODSWe collected secretory materials released from ASCs that had been transfected with antifibrotic miR-122 (MCM) and compared their antifibrotic effects with those of the naïve secretome (CM). MCM and CM were intravenously administered to the mouse model of thioacetamide-induced liver fibrosis, and their therapeutic potentials were compared.RESULTSMCM infusion provided higher therapeutic potential in terms of: (A) Reducing collagen content in the liver; (B) Inhibiting proinflammatory cytokines; and (C) Reducing abnormally elevated liver enzymes than the infusion of the naïve secretome. The proteomic analysis of MCM also indicated that the contents of antifibrotic proteins were significantly elevated compared to those in the naïve secretome.CONCLUSIONWe could, thus, conclude that the secretome released from miR-122-transfected ASCs has higher antifibrotic and anti-inflammatory properties than the naïve secretome. Because miR-122 transfection into ASCs provides a specific way of potentiating the antifibrotic properties of ASC secretome, it could be considered as an enhanced method for reinforcing secretome effectiveness.

A novel strategy to promote liver regeneration: utilization of secretome obtained from survivin-overexpressing adipose-derived stem cells

“…ASCs were plated into culture flask in Dulbecco’s Modified Eagle Medium with low glucose (DMEM/low glucose; Hyclone, Logan, UT, USA) supplemented with 0.1 mg/mL of streptomycin (Gibco BRL, Carlsbad, CA, USA), 100 U/mL of penicillin (Gibco), 10% fetal bovine serum (FBS; Hyclone) at 37°C in humidified chamber containing 5% carbon dioxide [ 8 12 ].…”

“…After 24 hours, ASCs were refed serum-free DMEM/low glucose for 24 hours. Then, the secretome was obtained 200 µL amount at ASCs, the CM were concentrated by 25 fold, by ultrafiltration device with a 3-kDa cutoff (Amicon Ultra-15; Millipore, Bedford, MA, USA) [ 8 ].…”

“…Recently, the use of stem cell secretome instead of stem cells has been actively studied in an attempt to overcome these limitations [ 6 ] secretome refers to substances secreted by the stem cells into the surrounding extracellular space [ 7 ]. The use of the secretome is supported by evidence that they mediate the main mechanism of action of stem cells [ 8 ]. Conditioned media (CM) obtained from MSCs were found to contain high levels of paracrine cytokines and growth factors that promote tissue repair in numerous in vitro and in vivo experimental models [ 6 ].…”

Applications of Mesenchymal Stem Cells in Liver Fibrosis: Novel Strategies, Mechanisms, and Clinical Practice

“…Lou et al showed that miR-122 modified adipose tissue-derived MSCs (AMSC-122), which were constructed through lentivirus-mediated transfer of pre-miR-122, more effectively inhibited the proliferation of hepatic stellate cells (HSCs) and the maturation of collagen [45]. In the latest study, the secreted proteome released from AMSC-122 has higher antifibrotic and antiinflammatory properties than the pure secreted proteome [46]. The research by Chen et al reported that inhibition of miR-26a-5p can further improve the therapeutic effect of MSCs on liver cirrhosis through increasing the expression of HGF protein in MSCs [47].…”

Circular RNA cMTO1 Promotes PTEN Expression Through Sponging miR-181b-5p in Liver Fibrosis

“…miRNAs have been reported to be deregulated in various human diseases and involved in many biological processes such as cell proliferation ( Ma et al, 2018 ; Bruch et al, 2019 ). Currently, a growing body of evidence suggests that miRNAs participate in the control of hepatic stellate cell (HSC) activation, which is an essential event in liver fibrosis ( Kim et al, 2019 ). We previously found that miR-17-5p, up-regulated in activated HSCs, promotes HSC activation via targeting of smad7 ( Yu et al, 2015 ).…”