Intrinsic Type 1 Interferon (IFN1) Profile of Uncultured Human Bone Marrow CD45lowCD271+ Multipotential Stromal Cells (BM-MSCs): The Impact of Donor Age, Culture Expansion and IFNα and IFNβ Stimulation

Abstract

Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence. Considering that BM-HSCs and BM-MSCs share the same BM niche, we investigated IFN1 expression profile in human BM-MSCs in relation to donor age, culture-expansion and IFN1 (α and β) stimulation. Fluorescence-activated cell sorting was used to purify uncultured BM-MSCs from younger (19–41, n = 6) and older (59–89, n = 6) donors based on the CD45lowCD271+ phenotype, and hematopoietic-lineage cells (BM-HLCs, CD45+CD271−) were used as controls. Gene expression was analysed using integrated circuits arrays in sorted fractions as well as cultured/stimulated BM-MSCs and Y201/Y202 immortalised cell lines. IFN1 stimulation led to BM-MSC growth arrest and upregulation of many IFN1-stimulated genes (ISGs), with IFNβ demonstrating stronger effects. Uncultured MSCs were characterised by a moderate-level ISG expression similar to Y201 cells. Age-related changes in ISG expression were negligible in BM-MSCs compared to BM-HLCs, and intracellular reactive oxygen species (ROS) levels in BM-MSCs did not significantly correlate with donor age. Antiaging genes Klotho and SIRT6 correlated with more ISGs in BM-MSCs than in BM-HLCs. In patients with osteoarthritis (OA), BM-MSCs expressed considerably lower levels of several ISGs, indicating that their IFN1 signature is affected in a pathological condition. In summary, BM-MSCs possess homeostatic IFN1 gene expression signature in health, which is sensitive to in vitro culture and external IFN1 stimulation. IFN signalling may facilitate in vivo BM-MSC responses to DNA damage and combating senescence and aberrant immune activation.

Multipotent Mesenchymal Stromal Cells in Rheumatoid Arthritis and Systemic Lupus Erythematosus; From a Leading Role in Pathogenesis to Potential Therapeutic Saviors?

“…With regards to accessibility, UC is a non-invasive source of MSCs, while harvesting BM-MSCs or AD-MSCs would require a surgical procedure, could sometimes be hard to access and cause complications ( 204 ). Additionally, MSC yield and functional potential are dependent on donor factors (e.g., age and co-morbidity) ( 89 , 205 ). Several in vitro , preclinical and clinical studies remain inconclusive but suggest that the immunosuppressive function of UC-MSCs and AD-MSCs could be more potent than that of BM-MSCs ( 137 , 139 , 145 , 206 ).…”

“…In contrast to hematopoietic cells, IFN-α seems to have less effect on MSCs. The gene expression levels of differentiation markers of native MSCs were found unaffected by IFN-α ( 89 ). However, when MSCs were treated with IFN-α within an in vitro culture condition, inhibition of cell proliferation was reported ( 89 , 90 ).…”

“…The gene expression levels of differentiation markers of native MSCs were found unaffected by IFN-α ( 89 ). However, when MSCs were treated with IFN-α within an in vitro culture condition, inhibition of cell proliferation was reported ( 89 , 90 ). Additionally, the osteogenic differentiation potential of culture expanded MSCs was found to be suppressed by exposure to IFN-α ( 91 ).…”

Bone Marrow Multipotent Mesenchymal Stromal Cells as Autologous Therapy for Osteonecrosis: Effects of Age and Underlying Causes

“…Using uncultured MSCs, our group has recently shown no significant age-related increase in adipogenic or osteogenic differentiation transcripts or ROS levels in uncultured MSCs from old donors compared to young donors [ 73 , 80 ]. This contrasts with the higher ROS levels found in minimally cultured MSCs from older donors [ 81 ].…”

Aging, Bone Marrow and Next-Generation Sequencing (NGS): Recent Advances and Future Perspectives

“…Studies have reported a decline in their number [48,49], decline in their proliferative potential [50,51], increased bias towards adipogenesis at the expense of osteoblast formation, resulting in decline in bone formation, and enhanced fat formation [52] and decrease in migratory properties [53]. More recently, potential links have been made between the intrinsic type 1 interferon profile within MSCs and their ability to respond to DNA damage [54]. The work by Josephson et al established the effect of age-related inflammation in MSCs by demonstrating a decline in the number of MSCs and impaired bone regeneration capacity in mice models.…”