Advertisement

Characterizing the In Vivo Role of Candidate Leukemia Stem Cell Genes

Protocol
First Online:
  • 452 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2185)

Abstract

Acute myeloid leukemia (AML) is a disease caused by multiple distinct genomic events in the hematopoietic stem cell and progenitor compartment. To gain insight into the link between genetic mutations in AML and their clinical significance, AML mouse models are often employed. However, the breeding of genetically modified mouse models is a resource-intensive and time-consuming endeavor. Here, we describe a viral-based protocol to study the role of candidate leukemia stem cell (LSC) genes. Transplantation of virally transduced oncogenic drivers for AML with virally altered expression of candidate leukemia associated genes in murine primary bone marrow cells, is an effective alternative method to assess the impact of cooperating mutations in AML.

Key words

Hematopoietic stem cells Leukemia stem cells Acute myeloid leukemia Transplantation Retroviral transduction Lentiviral transduction 
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Shlush LI, Zandi S, Mitchell A et al (2014) Identification of pre-leukaemic haematopoietic stem cells in acute leukaemia. Nature 506:328CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bonnet D, Dick JE (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 3:730–737.  https://doi.org/10.1038/nm0797-730CrossRefGoogle Scholar
  3. 3.
    Eppert K, Takenaka K, Lechman ER et al (2011) Stem cell gene expression programs influence clinical outcome in human leukemia. Nat Med 17:1086CrossRefPubMedGoogle Scholar
  4. 4.
    Ng SWK, Mitchell A, Kennedy JA et al (2016) A 17-gene stemness score for rapid determination of risk in acute leukaemia. Nature 540:433–437.  https://doi.org/10.1038/nature20598CrossRefPubMedGoogle Scholar
  5. 5.
    Costello RT, Mallet F, Gaugler B et al (2000) Human acute myeloid leukemia CD34+/CD38- progenitor cells have decreased sensitivity to chemotherapy and Fas-induced apoptosis, reduced immunogenicity, and impaired dendritic cell transformation capacities. Cancer Res 60:4403–4411PubMedGoogle Scholar
  6. 6.
    Ishikawa F, Yoshida S, Saito Y et al (2007) Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region. Nat Biotechnol 25:1315–1321.  https://doi.org/10.1038/nbt1350CrossRefPubMedGoogle Scholar
  7. 7.
    Boyd AL, Aslostovar L, Reid J et al (2018) Identification of chemotherapy-induced leukemic-regenerating cells reveals a transient vulnerability of human AML recurrence. Cancer Cell 34:483–498.e5.  https://doi.org/10.1016/j.ccell.2018.08.007CrossRefPubMedGoogle Scholar
  8. 8.
    Gregory TK, Wald D, Chen Y et al (2009) Molecular prognostic markers for adult acute myeloid leukemia with normal cytogenetics. J Hematol Oncol 2:23.  https://doi.org/10.1186/1756-8722-2-23CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Ley TJ, Miller C, Ding L et al (2013) Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N Engl J Med 368:2059–2074.  https://doi.org/10.1056/NEJMoa1301689CrossRefPubMedGoogle Scholar
  10. 10.
    White BS, DiPersio JF (2014) Genomic tools in acute myeloid leukemia: from the bench to the bedside. Cancer 120:1134–1144.  https://doi.org/10.1002/cncr.28552CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Welch JS, Ley TJ, Link DC et al (2012) The origin and evolution of mutations in acute myeloid leukemia. Cell 150:264–278.  https://doi.org/10.1016/j.cell.2012.06.023CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Woiterski J, Ebinger M, Witte KE et al (2013) Engraftment of low numbers of pediatric acute lymphoid and myeloid leukemias into NOD/SCID/IL2Rcgammanull mice reflects individual leukemogenecity and highly correlates with clinical outcome. Int J Cancer 133:1547–1556.  https://doi.org/10.1002/ijc.28170CrossRefGoogle Scholar
  13. 13.
    Pabst C, Bergeron A, Lavallee V-P et al (2016) GPR56 identifies primary human acute myeloid leukemia cells with high repopulating potential in vivo. Blood 127:2018–2027.  https://doi.org/10.1182/blood-2015-11-683649CrossRefPubMedGoogle Scholar
  14. 14.
    Cabezas-Wallscheid N, Eichwald V, de Graaf J et al (2013) Instruction of haematopoietic lineage choices, evolution of transcriptional landscapes and cancer stem cell hierarchies derived from an AML1-ETO mouse model. EMBO Mol Med 5:1804–1820.  https://doi.org/10.1002/emmm.201302661CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Lehnertz B, Zhang YW, Boivin I et al (2017) H3(K27M/I) mutations promote context-dependent transformation in acute myeloid leukemia with RUNX1 alterations. Blood 130:2204–2214.  https://doi.org/10.1182/blood-2017-03-774653CrossRefPubMedGoogle Scholar
  16. 16.
    Whitt MA (2010) Generation of VSV pseudotypes using recombinant DeltaG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses to vaccines. J Virol Methods 169:365–374.  https://doi.org/10.1016/j.jviromet.2010.08.006CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Zhang YW, Cabezas-Wallscheid N (2019) Assessment of young and aged hematopoietic stem cell activity by competitive serial transplantation assays. Methods Mol Biol 2017:193–203.  https://doi.org/10.1007/978-1-4939-9574-5_15CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Max Planck Institute of Immunobiology and EpigeneticsFreiburgGermany
  2. 2.International Max Planck Research School for Molecular and Cellular Biology (IMPRS-MCB)FreiburgGermany
  3. 3.Spemann Graduate School for Biology and Medicine (SGBM)FreiburgGermany
  4. 4.Centre for Integrative Biological Signalling Studies (CIBSS)FreiburgGermany

Personalised recommendations

Cite protocol

Buy options