Exofucosylation of Adipose Mesenchymal Stromal Cells Alters Their Secretome Profile

Abstract

Mesenchymal stromal cells (MSCs) constitute the cell type more frequently used in many regenerative medicine approaches due to their exclusive immunomodulatory properties, and they have been reported to mediate profound immunomodulatory effects in vivo. Nevertheless, MSCs do not express essential adhesion molecules actively involved in cell migration, a phenotypic feature that hampers their ability to home inflamed tissues following intravenous administration. In this study, we investigated whether modification by fucosylation of murine AdMSCs (mAdMSCs) creates Hematopoietic Cell E-/L-selectin Ligand, the E-selectin-binding CD44 glycoform. This cell surface glycan modification of CD44 has previously shown in preclinical studies to favor trafficking of mAdMSCs to inflamed or injured peripheral tissues. We analyzed the impact that exofucosylation could have in other innate phenotypic and functional properties of MSCs. Compared to unmodified counterparts, fucosylated mAdMSCs demonstrated higher in vitro migration, an altered secretome pattern, including increased expression and secretion of anti-inflammatory molecules, and a higher capacity to inhibit mitogen-stimulated splenocyte proliferation under standard culture conditions. Together, these findings indicate that exofucosylation could represent a suitable cell engineering strategy, not only to facilitate the in vivo MSC colonization of damaged tissues after systemic administration, but also to convert MSCs in a more potent immunomodulatory/anti-inflammatory cell therapy-based product for the treatment of a variety of autoimmune, inflammatory, and degenerative diseases.

Neuroprotection and Axonal Regeneration Induced by Bone Marrow Mesenchymal Stromal Cells Depend on the Type of Transplant

“…When cultures were 70–80% confluent, human and mouse BM-MSCs were subcultured at 5 × 10 3 cells/cm 2 and used in passages 3–4 for subsequent experiments. Human and mouse BM-MSCs were immunophenotypically characterized by flow cytometry (FACS Canto II, BD Biosciences, San Jose, CA, United States) as previously described ( Millán-Rivero et al, 2019 ; García-Bernal et al, 2020 ).…”

“…Human and mouse BM-MSCs were immunophenotypically characterized by flow cytometry (FACS Canto II, BD Biosciences, San Jose, CA, United States) as previously described (Millán-Rivero et al, 2019;García-Bernal et al, 2020).…”

The Current Status of Mesenchymal Stromal Cells: Controversies, Unresolved Issues and Some Promising Solutions to Improve Their Therapeutic Efficacy

“…This transient modification increases efficiently the in vivo tethering and rolling contacts on E-selectin-expressing endothelial beds in bone marrow microvasculature and in inflamed tissues ( Sackstein et al, 2008 ; Abdi et al, 2015 ; Dykstra et al, 2016 ; Chou et al, 2017 ). Remarkably, recent findings have shown that exofucosylated MSCs display an altered secretome characterized by an augmented expression of anti-inflammatory molecules, leading to higher MSC immunosuppressive properties, as well as increased migration ability toward some pro-inflammatory chemokines such as CCL5, CCL20 and CXCL16 ( Garcia-Bernal et al, 2020 ). Other authors reported that covalent binding of sLeX to the MSC surface through a biotin-streptavidin bridge, by conjugation of E-selectin-targeting peptide on the MSC membrane or by mRNA transfection to overexpress PSGL-1 and sLeX on MSCs resulted in an augmented rolling behavior on P- and E-selectin-coated surfaces and on inflamed vascular endothelium in vivo ( Sarkar et al, 2008 ; Cheng et al, 2012 ; Levy et al, 2013 ).…”

Mesenchymal stem cells for lysosomal storage and polyglutamine disorders: Possible shared mechanisms

“…Different medium composition and devices for cell growth (like bioreactors allowing for 3D cultures) as well as preconditioning methods, such as hypoxia, exposure to inflammatory factors or other chemicals, cell modifications (e.g. to overexpress certain molecules like growth factors or exo‐fucosylation – cell surface glycan modification of CD44) or even the use of MSC in combination with other therapies have been explored to increase cell viability, targeting and/or their therapeutic properties 31‐36 …”