Advertisement

Mesenchymal stem/stromal cells from a transplanted, asymptomatic patient with Fanconi anemia exhibit an aging-like phenotype and dysregulated expression of genes implicated in hematopoiesis and myelodysplasia,

Published:December 05, 2022DOI:https://doi.org/10.1016/j.jcyt.2022.11.003

      Abstract

      Background aims

      Fanconi anemia (FA) is an inherited bone marrow failure syndrome caused by defects in the repair of DNA inter-strand crosslinks and manifests as aplastic anemia, myelodysplastic syndrome and acute myeloid leukemia. FA also causes defects in mesenchymal stromal cell (MSC) function, but how different FA gene mutations alter function remains understudied.

      Methods

      We compared the growth, differentiation and transcript profile of a single MSC isolate from an asymptomatic patient with FA with a FANCG nonsense mutation who underwent hematopoietic stem cell transplantation 10 years prior to that from a representative healthy donor (HD).

      Results

      We show that FANCG−/− MSCs exhibit rapid onset of growth cessation, skewed bi-lineage differentiation in favor of adipogenesis and increased cellular oxidate stress consistent with an aging-like phenotype. Transcript profiling identified pathways related to cell growth, senescence, cellular stress responses and DNA replication/repair as over-represented in FANCG−/− MSC, and real-time polymerase chain reaction confirmed these MSCs expressed reduced levels of transcripts implicated in cell growth (TWIST1, FGFR2v7-8) and osteogenesis (TWIST1, RUNX2) and increased levels of transcripts regulating adipogenesis (GPR116) and insulin signaling. They also expressed reduced levels of mRNAs implicated in HSC self-maintenance and homing (KITLG, HGF, GDNF, PGF, CFB, IL-1B and CSF1) and elevated levels of those implicated in myelodysplasia (IL-6, GDF15).

      Conclusions

      Together, these findings demonstrate how inactivation of FANCG impacts MSC behavior, which parallels observed defects in osteogenesis, HSC depletion and leukemic blast formation seen in patients with FA.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Cytotherapy
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Díaz De Heredia C.
        • Bierings M.
        • Dalle J.H.
        • Fioredda F.
        • Strahm B.
        Fanconi's anemia and other hereditary bone marrow failure syndromes.
        in: Carreras E Dufour C Mohty M Kröger N The Ebmt Handbook: Hematopoietic Stem Cell Transplantation and Cellular Therapies. Springer, 2019: 587-593 (Copyright 2019, EBMT and the Author(s))
        • Ahmad S.I.
        • Hanaoka F.
        • Kirk S.H.
        Molecular biology of Fanconi anaemia–an old problem, a new insight.
        Bioessays. 2002; 24: 439-448
        • Shimamura A.
        • Alter B.P.
        Pathophysiology and management of inherited bone marrow failure syndromes.
        Blood Rev. 2010; 24: 101-122
        • Mathew C.G.
        Fanconi anaemia genes and susceptibility to cancer.
        Oncogene. 2006; 25: 5875-5884
        • Garaycoechea J.I.
        • Patel K.J.
        Why does the bone marrow fail in Fanconi anemia?.
        Blood. 2014; 123: 26-34
        • Kutler D.I.
        • et al.
        A 20-year perspective on the International Fanconi Anemia Registry (IFAR).
        Blood. 2003; 101: 1249-1256
        • Bhandari J.
        • Thada P.K.
        • Puckett Y.
        Fanconi anemia.
        Statpearls. StatPearls Publishing. StatPearls Publishing LLC., 2020 (Copyright © 2020)
        • Li Y.
        • Amarachintha S.
        • Wilson A.F.
        • Li X.
        • Du W.
        Persistent response of Fanconi anemia haematopoietic stem and progenitor cells to oxidative stress.
        Cell Cycle. 2017; 16: 1201-1209
        • Pagano G.
        • et al.
        Oxidative stress in Fanconi anaemia: from cells and molecules towards prospects in clinical management.
        Biological Chemistry. 2012; 393
        • Du W.
        • et al.
        The Fa pathway counteracts oxidative stress through selective protection of antioxidant defense gene promoters.
        Blood. 2012; 119: 4142-4151
        • Du W.
        • Adam Z.
        • Rani R.
        • Zhang X.
        • Pang Q.
        Oxidative stress in Fanconi anemia hematopoiesis and disease progression.
        Antioxid Redox Signal. 2008; 10: 1909-1921
        • Mukhopadhyay S.S.
        • et al.
        Defective mitochondrial peroxiredoxin-3 results in sensitivity to oxidative stress in Fanconi anemia.
        J Cell Biol. 2006; 175: 225-235
        • Ceccaldi R.
        • et al.
        Bone marrow failure in Fanconi anemia is triggered by an exacerbated P53/P21 DNA damage response that impairs hematopoietic stem and progenitor cells.
        Cell Stem Cell. 2012; 11: 36-49
        • Zhang H.
        • et al.
        TGF-β inhibition rescues hematopoietic stem cell defects and bone marrow failure in Fanconi anemia.
        Cell Stem Cell. 2016; 18: 668-681
        • Rodríguez A.
        • et al.
        MYC promotes bone marrow stem cell dysfunction in Fanconi anemia.
        Cell Stem Cell. 2021; 28: 33-47
        • Sacchetti B.
        • et al.
        Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment.
        Cell. 2007; 131: 324-336
        • Muguruma Y.
        • et al.
        Reconstitution of the functional human hematopoietic microenvironment derived from human mesenchymal stem cells in the murine bone marrow compartment.
        Blood. 2006; 107: 1878-1887
        • Majumdar M.K.
        • Thiede M.A.
        • Haynesworth S.E.
        • Bruder S.P.
        • Gerson S.L.
        Human marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineages.
        J Hematother Stem Cell Res. 2000; 9: 841-848
        • Li T.
        • Wu Y.
        Paracrine molecules of mesenchymal stem cells for hematopoietic stem cell niche.
        Bone Marrow Res. 2011; 2011353878
        • Deryugina E.I.
        • Muller-Sieburg C.E.
        Stromal Cells in Long-Term Cultures: Keys to the Elucidation of Hematopoietic Development?.
        Crit Rev Immunol. 1993; 13: 115-150
        • Zhang Q.S.
        • et al.
        Fancd2–/– mice have hematopoietic defects that can be partially corrected by resveratrol.
        Blood. 2010; 116: 5140-5148
        • Li Y.
        • et al.
        Mesenchymal stem/progenitor cells promote the reconstitution of exogenous hematopoietic stem cells in Fancg–/– mice in vivo.
        Blood. 2009; 113: 2342-2351
        • Pulliam-Leath A.C.
        • et al.
        Genetic disruption of both Fancc and Fancg in mice recapitulates the hematopoietic manifestations of Fanconi anemia.
        Blood. 2010; 116: 2915-2920
        • Lecourt S.
        • et al.
        Bone marrow microenvironment in fanconi anemia: a prospective functional study in a cohort of Fanconi anemia patients.
        Stem Cells Dev. 2010; 19: 203-208
        • Cagnan I.
        • et al.
        Bone marrow mesenchymal stem cells carrying fancd2 mutation differ from the other Fanconi anemia complementation groups in terms of TGF-β1 Production.
        Stem Cell Rev Rep. 2018; 14: 425-437
        • Mantelli M.
        • et al.
        Comprehensive characterization of mesenchymal stromal cells from patients with Fanconi anaemia.
        Br J Haematol. 2015; 170: 826-836
        • Russell K.C.
        • et al.
        In vitro high-capacity assay to quantify the clonal heterogeneity in trilineage potential of mesenchymal stem cells reveals a complex hierarchy of lineage commitment.
        Stem Cells. 2010; 28: 788-798
        • Boregowda S.V.
        • et al.
        Atmospheric oxygen inhibits growth and differentiation of marrow-derived mouse mesenchymal stem cells via a P53-dependent mechanism: implications for long-term culture expansion.
        Stem Cells. 2012; 30: 975-987
        • Coipeau P.
        • et al.
        Impaired differentiation potential of human trabecular bone mesenchymal stromal cells from elderly patients.
        Cytotherapy. 2009; 11: 584-594
        • Moerman E.J.
        • Teng K.
        • Lipschitz D.A.
        • Lecka-Czernik B.
        Aging activates adipogenic and suppresses osteogenic programs in mesenchymal marrow stroma/stem cells: the role of PPAR-gamma2 transcription factor and TGF-beta/bmp signaling pathways.
        Aging Cell. 2004; 3: 379-389
        • Pfützner A.
        • et al.
        Mesenchymal stem cell differentiation into adipocytes is equally induced by insulin and proinsulin in vitro.
        Int J Stem Cells. 2017; 10: 154-159
        • Paul J.D.
        • et al.
        Slit3-Robo4 activation promotes vascular network formation in human engineered tissue and angiogenesis in vivo.
        J Mol Cell Cardiol. 2013; 64: 124-131
        • Smith-Berdan S.
        • Schepers K.
        • Ly A.
        • Passegue E.
        • Forsberg E.C.
        Dynamic expression of the Robo ligand Slit2 in bone marrow cell populations.
        Cell Cycle. 2012; 11: 675-682
        • Sillat T.
        • et al.
        Basement membrane collagen type IV expression by human mesenchymal stem cells during adipogenic differentiation.
        J Cell Mol Med. 2012; 16: 1485-1495
        • Boregowda S.V.
        • Krishnappa V.
        • Haga C.L.
        • Ortiz L.A.
        • Phinney D.G.
        A clinical indications prediction scale based on Twist1 for human mesenchymal stem cells.
        EBioMedicine. 2016; 4: 62-73
        • Säily M.
        • Koistinen P.
        • Savolainen E.R.
        The soluble form of interleukin-6 receptor modulates cell proliferation by acute myeloblastic leukemia blast cells.
        Ann Hematol. 1999; 78: 173-179
        • Phinney D.G.
        • et al.
        Mesenchymal stem cells use extracellular vesicles to outsource mitophagy and shuttle MicroRNAs.
        Nat Commun. 2015; 6: 8472
        • Franceschi R.T.
        The developmental control of osteoblast-specific gene expression: role of specific transcription factors and the extracellular matrix environment.
        Crit Rev Oral Biol Med. 1999; 10: 40-57
        • Nie T.
        • et al.
        Adipose tissue deletion of Gpr116 impairs insulin sensitivity through modulation of adipose function.
        FEBS Lett. 2012; 586: 3618-3625
        • Schepers K.
        • Campbell T.B.
        • Passegue E.
        Normal and leukemic stem cell niches: insights and therapeutic opportunities.
        Cell Stem Cell. 2015; 16: 254-267
        • Koc O.N.
        • et al.
        Rapid hematopoietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patients receiving high-dose chemotherapy.
        J Clin Oncol. 2000; 18: 307-316
        • Desbourdes L.
        • et al.
        Alteration analysis of bone marrow mesenchymal stromal cells from de novo acute myeloid leukemia patients at diagnosis.
        Stem Cells Dev. 2017; 26: 709-722
        • Chandran P.
        • et al.
        Mesenchymal stromal cells from patients with acute myeloid leukemia have altered capacity to expand differentiated hematopoietic progenitors.
        Leuk Res. 2015; 39: 486-493
        • Geyh S.
        • et al.
        Insufficient stromal support in MDS results from molecular and functional deficits of mesenchymal stromal cells.
        Leukemia. 2013; 27: 1841-1851
        • Zhao Z.G.
        • et al.
        Functional characteristics of mesenchymal stem cells derived from bone marrow of patients with myelodysplastic syndromes.
        Cancer Lett. 2012; 317: 136-143
        • Fattizzo B.
        • Giannotta J.A.
        • Barcellini W.
        Mesenchymal stem cells in aplastic anemia and myelodysplastic syndromes: the "seed and soil" crosstalk.
        Int J Mol Sci. 2020; 21: 5438
        • Zambetti N.A.
        • et al.
        Mesenchymal inflammation drives genotoxic stress in hematopoietic stem cells and predicts disease evolution in human pre-leukemia.
        Cell Stem Cell. 2016; 19: 613-627
        • Zhai Y.
        • et al.
        Growth differentiation factor 15 contributes to cancer-associated fibroblasts-mediated chemo-protection of AML cells.
        J Exp Clin Cancer Res. 2016; 35: 147
        • Corre J.
        • et al.
        Bone marrow mesenchymal stem cells are abnormal in multiple myeloma.
        Leukemia. 2007; 21: 1079-1088
        • O'hagan-Wong K.
        • et al.
        Increased Il-6 secretion by aged human mesenchymal stromal cells disrupts hematopoietic stem and progenitor cells' homeostasis.
        Oncotarget. 2016; 7: 13285-13296
        • Gnani D.
        • et al.
        An early-senescence state in aged mesenchymal stromal cells contributes to hematopoietic stem and progenitor cell clonogenic impairment through the activation of a pro-inflammatory program.
        Aging Cell. 2019; 18: e12933
        • Bartsch K.
        • et al.
        Mesenchymal stem cells remain host-derived independent of the source of the stem-cell graft and conditioning regimen used.
        Transplantation. 2009; 87: 217-221
        • Rieger K.
        • et al.
        Mesenchymal STEM CELLS REMAIN OF HOST ORIGIN EVEN A LONG TIME AFTER ALLOGENEIC PERIPHERAL BLOOD STEM CELL OR BONE MARROW TRANSPLANTATION.
        Exp Hematol. 2005; 33: 605-611
        • Sanchez-Guijo F.M.
        • et al.
        Posttransplant hematopoiesis in patients undergoing sibling allogeneic stem cell transplantation reflects that of their respective donors although with a lower functional capability.
        Exp Hematol. 2005; 33: 935-943
        • Banfi A.
        • Bianchi G.
        • Galotto M.
        • Cancedda R.
        • Quarto R.
        Bone marrow stromal damage after chemo/radiotherapy: occurrence, consequences and possibilities of treatment.
        Leuk Lymphoma. 2001; 42: 863-870
        • O'flaherty E.
        • Sparrow R.
        • Szer J.
        Bone marrow stromal function from patients after bone marrow transplantation.
        Bone Marrow Transplant. 1995; 15: 207-212
        • Anur P.
        • et al.
        Late effects in patients with Fanconi anemia following allogeneic hematopoietic stem cell transplantation from alternative donors.
        Bone Marrow Transplant. 2016; 51: 938-944