In this study we propose a novel approach based on the use of mesenchymal stromal cells (MSCs), aiming at limiting risks of graft failure in gene therapy protocols associated with low conditioning regimens. Because the engraftment of corrected hematopoietic stem cells (HSCs) is particularly challenging in Fanconi anemia (FA), we have investigated the relevance of MSCs in an experimental model of FA gene therapy. Our results showed, first, that risks of graft failure in recipients conditioned with a moderate dose of 5 Gy and infused with limited numbers of wild-type HSCs are significantly higher in Fanca−/− recipients as compared with wild-type recipients. However, when wild-type HSC numbers inducing 30–50% of graft failures in Fanca−/− recipients were coinfused with MSCs, no graft failures were observed. Moreover, graft failures associated with the infusion of low numbers of gene-corrected Fanca−/− HSCs were also significantly overcome by MSC coinfusion. Our study shows for the first time that MSC coinfusion constitutes a simple and nontoxic approach to minimize risks of graft failure in gene therapy applications associated with low conditioning regimens and infusion of limited numbers of corrected HSCs.
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References
1.
NaldiniL. Ex vivo gene transfer and correction for cell-based therapies. Nat Rev Genet, 2015; 12:301–315.
2.
BernardoME, AiutiA. The role of conditioning in hematopoietic stem cell gene therapy. Hum Gene Ther, 2016:27:741–748.
3.
FriedensteinAJ, LatzinikNW, GroshevaAG, et al.Marrow microenvironment transfer by heterotopic transplantation of freshly isolated and cultured cells in porous sponges. Exp Hematol, 1982; 10:217–227.
4.
PittengerMF, MackayAM, BeckSC, et al.Multilineage potential of adult human mesenchymal stem cells. Science, 1999; 284:143–147.
5.
CongetPA, MinguellJJ. Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol, 1999; 181:67–73.
6.
Le BlancK. Immunomodulatory effects of fetal and adult mesenchymal stem cells. Cytotherapy, 2003; 5:485–489.
7.
YanezR, OviedoA, AldeaM, et al.Prostaglandin E2 plays a key role in the immunosuppressive properties of adipose and bone marrow tissue-derived mesenchymal stromal cells. Exp Cell Res, 2010; 316:3109–3123.
8.
BernardoME, FibbeWE. Mesenchymal stromal cells: sensors and switchers of inflammation. Cell Stem Cell, 2013; 13:392–402.
9.
CarrancioS, RomoC, RamosT, et al.Effects of MSC coadministration and route of delivery on cord blood hematopoietic stem cell engraftment. Cell Transplant, 2012; 22:1171–1183.
10.
KimDH, YooKH, YimYS, et al.Cotransplanted bone marrow derived mesenchymal stem cells (MSC) enhanced engraftment of hematopoietic stem cells in a MSC-dose dependent manner in NOD/SCID mice. J Korean Med Sci, 2006; 21:1000–1004.
11.
AngelopoulouM, NovelliE, GroveJE, et al.Cotransplantation of human mesenchymal stem cells enhances human myelopoiesis and megakaryocytopoiesis in NOD/SCID mice. Exp Hematol, 2003; 31:413–420.
12.
ParkSK, WonJH, KimHJ, et al.Co-transplantation of human mesenchymal stem cells promotes human CD34+ cells engraftment in a dose-dependent fashion in NOD/SCID mice. J Korean Med Sci, 2007; 22:412–419.
13.
NoortWA, KruisselbrinkAB, in't AnkerPS, et al.Mesenchymal stem cells promote engraftment of human umbilical cord blood-derived CD34+ cells in NOD/SCID mice. Exp Hematol, 2002; 30:870–878.
14.
Le BlancK, SamuelssonH, GustafssonB, et al.Transplantation of mesenchymal stem cells to enhance engraftment of hematopoietic stem cells. Leukemia, 2007; 21:1733–1738.
15.
BallLM, BernardoME, RoelofsH, et al.Cotransplantation of ex vivo expanded mesenchymal stem cells accelerates lymphocyte recovery and may reduce the risk of graft failure in haploidentical hematopoietic stem-cell transplantation. Blood, 2007; 110:2764–2767.
16.
FangB, LiN, SongY, et al.Cotransplantation of haploidentical mesenchymal stem cells to enhance engraftment of hematopoietic stem cells and to reduce the risk of graft failure in two children with severe aplastic anemia. Pediatr Transplant, 2009; 13:499–502.
17.
Fernandez-GarciaM, YanezRM, Sanchez-DominguezR, et al.Mesenchymal stromal cells enhance the engraftment of hematopoietic stem cells in an autologous mouse transplantation model. Stem Cell Res Ther, 2015; 6:165.
18.
KellyPF, RadtkeS, KalleC, et al.Stem cell collection and gene transfer in Fanconi anemia. Mol Ther, 2007; 15:211–219.
19.
LiuJM, KimS, ReadEJ, et al.Engraftment of hematopoietic progenitor cells transduced with the Fanconi anemia group C gene (FANCC). Hum Gene Ther, 1999; 10:2337–2346.
20.
CroopJM, CooperR, FernandezC, et al.Mobilization and collection of peripheral blood CD34+ cells from patients with Fanconi anemia. Blood, 2001; 98:2917–2921.
21.
CasadoJA, CallenE, JacomeA, et al.A comprehensive strategy for the subtyping of Fanconi anemia patients: conclusions from the Spanish Fanconi Anemia Research Network. J Med Genet, 2007; 44:241–249.
22.
DongH, NebertDW, BrufordEA, et al.Update of the human and mouse Fanconi anemia genes. Hum Genomics, 2015; 9:32.
23.
KottemannMC, SmogorzewskaA. Fanconi anaemia and the repair of Watson and Crick DNA crosslinks. Nature, 2013; 493:356–363.
24.
MichlJ, ZimmerJ, TarsounasM. Interplay between Fanconi anemia and homologous recombination pathways in genome integrity. EMBO J, 2016; 35:909–923.
25.
ChengNC, van de VrugtHJ, van der ValkMA, et al.Mice with a targeted disruption of the Fanconi anemia homolog Fanca. Hum Mol Genet, 2000; 9:1805–1811.
26.
Gonzalez-MurilloA, LozanoML, AlvarezL, et al.Development of lentiviral vectors with optimized transcriptional activity for the gene therapy of patients with Fanconi anemia. Hum Gene Ther, 2010; 21:623–630.
27.
HuY, SmythGK. ELDA: extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J Immunol Methods, 2009; 347:70–78.
28.
CarreauM, GanOI, LiuL, et al.Hematopoietic compartment of Fanconi anemia group C null mice contains fewer lineage-negative CD34+ primitive hematopoietic cells and shows reduced reconstruction ability. Exp Hematol, 1999; 27:1667–1674.
29.
LiX, PlettPA, YangY, et al.Fanconi anemia type C-deficient hematopoietic stem/progenitor cells exhibit aberrant cell cycle control. Blood, 2003; 102:2081–2084.
30.
PalchaudhuriR, SaezB, HoggattJ, et al.Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol, 2016; 34:738–745.
31.
ChhabraA, RingAM, WeiskopfK, et al.Hematopoietic stem cell transplantation in immunocompetent hosts without radiation or chemotherapy. Sci Transl Med, 2016; 8:351ra105.
32.
ChengNC, van de VrugtHJ, van der ValkMA, et al.Mice with a targeted disruption of the Fanconi anemia homolog Fanca. Hum Mol Genet, 2000; 9:1805–1811.
33.
RioP, SegoviaJC, HanenbergH, et al.In vitro phenotypic correction of hematopoietic progenitors from Fanconi anemia group A knockout mice. Blood, 2002; 100:2032–2039.
34.
LiY, ChenS, YuanJ, et al.Mesenchymal stem/progenitor cells promote the reconstitution of exogenous hematopoietic stem cells in Fancg−/− mice in vivo. Blood, 2009; 113:2342–2351.
35.
GrossM, HanenbergH, LobitzS, et al.Reverse mosaicism in Fanconi anemia: natural gene therapy via molecular self-correction. Cytogenet Genome Res, 2002; 98:126–135.
36.
Molina-EstevezFJ, NowrouziA, LozanoML, et al.Lentiviral-mediated gene therapy in Fanconi anemia-A mice reveals long-term engraftment and continuous turnover of corrected HSCs. Curr Gene Ther, 2015; 15:550–562.
37.
DufourC, CorcioneA, SvahnJ, et al.TNF-α and IFN-γ are overexpressed in the bone marrow of Fanconi anemia patients and TNF-α suppresses erythropoiesis in vitro. Blood, 2003; 102:2053–2059.
38.
KramperaM, CosmiL, AngeliR, et al.Role for interferon-γ in the immunomodulatory activity of human bone marrow mesenchymal stem cells. Stem Cells, 2006; 24:386–398.
39.
RenG, ZhangL, ZhaoX, et al.Mesenchymal stem cell-mediated immunosuppression occurs via concerted action of chemokines and nitric oxide. Cell Stem Cell, 2008; 2:141–150.
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