Abstract
Although NOD-SCID IL2Rγ−/− (NSG) xenograft mice are currently the most frequently used model to study human leukemia in vivo, the absence of a human niche severely hampers faithful recapitulation of the disease. We used NSG mice in which ceramic scaffolds seeded with human mesenchymal stromal cells were implanted to generate a human bone marrow (huBM-sc)-like niche. We observed that, in contrast to the murine bone marrow (mBM) niche, the expression of BCR-ABL or MLL-AF9 was sufficient to induce both primary acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL). Stemness was preserved within the human niches as demonstrated by serial transplantation assays. Efficient engraftment of AML MLL-AF9 and blast-crisis chronic myeloid leukemia patient cells was also observed, whereby the immature blast-like phenotype was maintained in the huBM-sc niche but to a much lesser extent in mBM niches. We compared transcriptomes of leukemias derived from mBM niches versus leukemias from huBM-like scaffold-based niches, which revealed striking differences in the expression of genes associated with hypoxia, mitochondria and metabolism. Finally, we utilized the huBM-sc MLL-AF9 B-ALL model to evaluate the efficacy of the I-BET151 inhibitor in vivo. In conclusion, we have established human niche models in which the myeloid and lymphoid features of BCR-ABL+ and MLL-AF9+ leukemias can be studied in detail.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Agliano A, Martin-Padura I, Mancuso P, Marighetti P, Rabascio C, Pruneri G et al. Human acute leukemia cells injected in NOD/LtSz-scid/IL-2Rgamma null mice generate a faster and more efficient disease compared to other NOD/scid-related strains. Int J Cancer 2008; 123: 2222–2227.
Sanchez PV, Perry RL, Sarry JE, Perl AE, Murphy K, Swider CR et al. A robust xenotransplantation model for acute myeloid leukemia. Leukemia 2009; 23: 2109–2117.
Goyama S, Wunderlich M, Mulloy JC . Xenograft models for normal and malignant stem cells. Blood 2015; 125: 2630–2640.
Vargaftig J, Taussig DC, Griessinger E, njos-Afonso F, Lister TA, Cavenagh J et al. Frequency of leukemic initiating cells does not depend on the xenotransplantation model used. Leukemia 2011; 26: 858–860.
Horton SJ, Jaques J, Woolthuis C, van DJ, Mesuraca M, Huls G et al. MLL-AF9-mediated immortalization of human hematopoietic cells along different lineages changes during ontogeny. Leukemia 2012; 5: 1116–1126.
Rizo A, Horton SJ, Olthof S, Dontje B, Ausema A, van Os R et al. BMI1 collaborates with BCR-ABL in leukemic transformation of human CD34+ cells. Blood 2010; 116: 4621–4630.
Groen RW, Noort WA, Raymakers RA, Prins HJ, Aalders L, Hofhuis FM et al. Reconstructing the human hematopoietic niche in immunodeficient mice: opportunities for studying primary multiple myeloma. Blood 2012; 120: e9–e16.
Gutierrez A, Pan L, Groen RW, Baleydier F, Kentsis A, Marineau J et al. Phenothiazines induce PP2A-mediated apoptosis in T cell acute lymphoblastic leukemia. J Clin Invest 2014; 124: 644–655.
Schuringa JJ, Chung KY, Morrone G, Moore MA . Constitutive activation of STAT5A promotes human hematopoietic stem cell self-renewal and erythroid differentiation. J Exp Med 2004; 200: 623–635.
Chiarella E, Carra G, Scicchitano S, Codispoti B, Mega T, Lupia M et al. UMG Lenti: novel lentiviral vectors for efficient transgene- and reporter gene expression in human early hematopoietic progenitors. PLoS One 2014; 9: e114795.
Barabe F, Kennedy JA, Hope KJ, Dick JE . Modeling the initiation and progression of human acute leukemia in mice. Science 2007; 316: 600–604.
Dawson MA, Prinjha RK, Dittman A, Giotopoulos G, Bantscheff M, Chan WI et al. Inhibition of BET recruitment to chromatin as an effective treatment for MLL-fusion leukaemia. Nature 2011; 478: 529–533.
Fong CY, Gilan O, Lam EY, Rubin AF, Ftouni S, Tyler D et al. BET inhibitor resistance emerges from leukaemia stem cells. Nature 2015; 525: 529–542.
Wei J, Wunderlich M, Fox C, Alvarez S, Cigudosa JC, Wilhelm JS et al. Microenvironment determines lineage fate in a human model of MLL-AF9 leukemia. Cancer Cell 2008; 13: 483–495.
Theocharides AP, Rongvaux A, Fritsch K, Flavell RA, Manz MG . Humanized hemato-lymphoid system mice. Haematologica 2016; 101: 5–19.
Nicolini FE, Cashman JD, Hogge DE, Humphries RK, Eaves CJ . NOD/SCID mice engineered to express human IL-3, GM-CSF and Steel factor constitutively mobilize engrafted human progenitors and compromise human stem cell regeneration. Leukemia 2004; 18: 341–347.
Feuring-Buske M, Gerhard B, Cashman J, Humphries RK, Eaves CJ, Hogge DE . Improved engraftment of human acute myeloid leukemia progenitor cells in beta 2-microglobulin-deficient NOD/SCID mice and in NOD/SCID mice transgenic for human growth factors. Leukemia 2003; 17: 760–763.
Wunderlich M, Chou FS, Link KA, Mizukawa B, Perry RL, Carroll M et al. AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3. Leukemia 2010; 24: 1785–1788.
Rongvaux A, Willinger T, Martinek J, Strowig T, Gearty SV, Teichmann LL et al. Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 2014; 32: 364–372.
Rongvaux A, Willinger T, Takizawa H, Rathinam C, Auerbach W, Murphy AJ et al. Human thrombopoietin knockin mice efficiently support human hematopoiesis in vivo. Proc Natl Acad Sci USA 2011; 108: 2378–2383.
Strowig T, Rongvaux A, Rathinam C, Takizawa H, Borsotti C, Philbrick W et al. Transgenic expression of human signal regulatory protein alpha in Rag2-/-gamma(c)-/- mice improves engraftment of human hematopoietic cells in humanized mice. Proc Natl Acad Sci USA 2011; 108: 13218–13223.
Willinger T, Rongvaux A, Takizawa H, Yancopoulos GD, Valenzuela DM, Murphy AJ et al. Human IL-3/GM-CSF knock-in mice support human alveolar macrophage development and human immune responses in the lung. Proc Natl Acad Sci USA 2011; 108: 2390–2395.
Willinger T, Rongvaux A, Strowig T, Manz MG, Flavell RA . Improving human hemato-lymphoid-system mice by cytokine knock-in gene replacement. Trends Immunol 2011; 32: 321–327.
Medyouf H, Mossner M, Jann JC, Nolte F, Raffel S, Herrmann C et al. Myelodysplastic cells in patients reprogram mesenchymal stromal cells to establish a transplantable stem cell niche disease unit. Cell Stem Cell 2014; 14: 824–837.
Rhyasen GW, Wunderlich M, Tohyama K, Garcia-Manero G, Mulloy JC, Starczynowski DT . An MDS xenograft model utilizing a patient-derived cell line. Leukemia 2014; 28: 1142–1145.
Chalandon Y, Jiang X, Christ O, Loutet S, Thanopoulou E, Eaves A et al. BCR-ABL-transduced human cord blood cells produce abnormal populations in immunodeficient mice. Leukemia 2005; 19: 442–448.
Sengupta A, Ficker AM, Dunn SK, Madhu M, Cancelas JA . Bmi1 reprograms CML B-lymphoid progenitors to become B-ALL-initiating cells. Blood 2012; 119: 494–502.
Heddleston JM, Li Z, Lathia JD, Bao S, Hjelmeland AB, Rich JN . Hypoxia inducible factors in cancer stem cells. Br J Cancer 2010; 102: 789–795.
Keith B, Simon MC . Hypoxia-inducible factors, stem cells, and cancer. Cell 2007; 129: 465–472.
Lee KE, Simon MC . From stem cells to cancer stem cells: HIF takes the stage. Curr Opin Cell Biol 2012; 24: 232–235.
Matsumoto A, Takeishi S, Kanie T, Susaki E, Onoyama I, Tateishi Y et al. p57 is required for quiescence and maintenance of adult hematopoietic stem cells. Cell Stem Cell 2011; 9: 262–271.
Scandura JM, Boccuni P, Massague J, Nimer SD . Transforming growth factor beta-induced cell cycle arrest of human hematopoietic cells requires p57KIP2 up-regulation. Proc Natl Acad Sci USA 2004; 101: 15231–15236.
Yang J, Chai L, Liu F, Fink LM, Lin P, Silberstein LE et al. Bmi-1 is a target gene for SALL4 in hematopoietic and leukemic cells. Proc Natl Acad Sci USA 2007; 104: 10494–10499.
Wang X, Venugopal C, Manoranjan B, McFarlane N, O'Farrell E, Nolte S et al. Sonic hedgehog regulates Bmi1 in human medulloblastoma brain tumor-initiating cells. Oncogene 2012; 31: 187–199.
Acknowledgements
We thank Professor JJ Erich and Dr A van Loon and colleagues (Departments of Obstetrics, University Medical Center Groningen and Martini Hospital Groningen) for collecting cord blood; Jeanet Dales for help with CB isolations; and Henk Moes, Roelof Jan van der Lei and Geert Mesander from Flow Cytometrics Facility for technical assistance with cell sorting. This work was supported by grants from the Dutch Cancer Society (2009-4411; VU2011-5127) and by the EU (ITN EuroCSC). I-BET151 was kindly provided by Nicholas Smithers (GSK R&D, UK).
Author contributions
PS and MC designed and conducted experiments, performed analysis and wrote the manuscript; JJ, AZBV, LLA, RWJG and JJS conducted experiments and analyzed data; HY and JDB contributed scaffold materials; ACMM, RWJG and EV analyzed data and participated in writing the manuscript; JJS designed experiments, performed analysis, wrote the manuscript and supervised the project.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Leukemia website
Rights and permissions
About this article
Cite this article
Sontakke, P., Carretta, M., Jaques, J. et al. Modeling BCR-ABL and MLL-AF9 leukemia in a human bone marrow-like scaffold-based xenograft model. Leukemia 30, 2064–2073 (2016). https://doi.org/10.1038/leu.2016.108
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/leu.2016.108
This article is cited by
-
HIF1/2-exerted control over glycolytic gene expression is not functionally relevant for glycolysis in human leukemic stem/progenitor cells
Cancer & Metabolism (2019)
-
MLLT3 governs human haematopoietic stem-cell self-renewal and engraftment
Nature (2019)
-
FLT3N676K drives acute myeloid leukemia in a xenograft model of KMT2A-MLLT3 leukemogenesis
Leukemia (2019)
-
From haematopoietic stem cells to complex differentiation landscapes
Nature (2018)
-
Generation and use of a humanized bone-marrow-ossicle niche for hematopoietic xenotransplantation into mice
Nature Protocols (2017)