Co-administration of human MSC overexpressing HIF-1α increases human CD34 + cell engraftment in vivo.

Autor: Preciado S; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain.; RETIC TerCel, ISCIII, Madrid, Spain.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain., Sirerol-Piquer MS; Departamento de Biología Celular, Biología Funcional y Antropología Física, University of Valencia, Burjassot, Spain.; Instituto de Biotecnología y Biomedicina (BioTecMed), University of Valencia, Burjassot, Spain.; RETIC TerCel, ISCIII, Madrid, Spain., Muntión S; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain.; RETIC TerCel, ISCIII, Madrid, Spain.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain., Osugui L; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain.; RETIC TerCel, ISCIII, Madrid, Spain.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain., Martí-Chillón GJ; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain.; RETIC TerCel, ISCIII, Madrid, Spain.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain., Navarro-Bailón A; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain.; RETIC TerCel, ISCIII, Madrid, Spain.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain., Sepúlveda P; Regenerative Medicine and Heart Transplantation Unit, Instituto de Investigación Sanitaria La Fe, Valencia, Spain.; RETIC TerCel, ISCIII, Madrid, Spain., Sánchez-Guijo F; Cell Therapy Unit, Hematology Department, University Hospital of Salamanca, IBSAL, University of Salamanca, Paseo de San Vicente 58-182, 37007, Salamanca, Spain. ferminsg@usal.es.; RETIC TerCel, ISCIII, Madrid, Spain. ferminsg@usal.es.; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Salamanca, Spain. ferminsg@usal.es.
Jazyk: angličtina
Zdroj: Stem cell research & therapy [Stem Cell Res Ther] 2021 Dec 07; Vol. 12 (1), pp. 601. Date of Electronic Publication: 2021 Dec 07.
DOI: 10.1186/s13287-021-02669-z
Abstrakt: Background: Poor graft function or graft failure after allogeneic stem cell transplantation is an unmet medical need, in which mesenchymal stromal cells (MSC) constitute an attractive potential therapeutic approach. Hypoxia-inducible factor-1α (HIF-1α) overexpression in MSC (HIF-MSC) potentiates the angiogenic and immunomodulatory properties of these cells, so we hypothesized that co-transplantation of MSC-HIF with CD34 + human cord blood cells would also enhance hematopoietic stem cell engraftment and function both in vitro and in vivo.
Methods: Human MSC were obtained from dental pulp. Lentiviral overexpression of HIF-1α was performed transducing cells with pWPI-green fluorescent protein (GFP) (MSC WT) or pWPI-HIF-1α-GFP (HIF-MSC) expression vectors. Human cord blood CD34 + cells were co-cultured with MSC WT or HIF-MSC (4:1) for 72 h. Then, viability (Annexin V and 7-AAD), cell cycle, ROS expression and immunophenotyping of key molecules involved in engraftment (CXCR4, CD34, ITGA4, c-KIT) were evaluated by flow cytometry in CD34 + cells. In addition, CD34 + cells clonal expansion was analyzed by clonogenic assays. Finally, in vivo engraftment was measured by flow cytometry 4-weeks after CD34 + cell transplantation with or without intrabone MSC WT or HIF-MSC in NOD/SCID mice.
Results: We did not observe significant differences in viability, cell cycle and ROS expression between CD34 + cells co-cultured with MSC WT or HIF-MSC. Nevertheless, a significant increase in CD34, CXCR4 and ITGA4 expression (p = 0.009; p = 0.001; p = 0.013, respectively) was observed in CD34 + cells co-cultured with HIF-MSC compared to MSC WT. In addition, CD34 + cells cultured with HIF-MSC displayed a higher CFU-GM clonogenic potential than those cultured with MSC WT (p = 0.048). We also observed a significant increase in CD34 + cells engraftment ability when they were co-transplanted with HIF-MSC compared to CD34 + co-transplanted with MSC WT (p = 0.016) or alone (p = 0.015) in both the injected and contralateral femurs (p = 0.024, p = 0.008 respectively).
Conclusions: Co-transplantation of human CD34 + cells with HIF-MSC enhances cell engraftment in vivo. This is probably due to the ability of HIF-MSC to increase clonogenic capacity of hematopoietic cells and to induce the expression of adhesion molecules involved in graft survival in the hematopoietic niche.
(© 2021. The Author(s).)
Databáze: MEDLINE
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