CD69 marks a subpopulation of acute myeloid leukemia with enhanced colony forming capacity and a unique signaling activation state

Abstract

In acute myeloid leukemia (AML), leukemia stem cells (LSCs) have self-renewal potential and are responsible for relapse. We previously showed that, in Mll-AF9/NRAS^G12V murine AML, CD69 expression marks an LSC-enriched subpopulation with enhanced in vivo self-renewal capacity. Here, we used CyTOF to define activated signaling pathways in LSC subpopulations in Mll-AF9/NRAS^G12V AML. Furthermore, we compared the signaling activation states of CD69^High and CD36^High subsets of primary human AML. The human CD69^High subset expresses low levels of Ki67 and high levels of NFκB and pMAPKAPKII. Additionally, the human CD69^High AML subset also has enhanced colony-forming capacity. We applied Bayesian network modeling to compare the global signaling network within the human AML subsets. We find that distinct signaling states, distinguished by NFκB and pMAPKAPKII levels, correlate with divergent functional subsets, defined by CD69 and CD36 expression, in human AML. Targeting NFκB with proteasome inhibition diminished colony formation.

Highlights

1. Immunophenotypically-defined murine AML stem cells harbor self-renewing and non-self-renewing subsets that display unique signaling characteristics.

2. CD69, an NFκB target gene, marks a subset of human AML with increased colony forming capacity and reduced proliferation.

3. NFκB activation correlates with the global signaling pathway activation state in human AML.

Keywords:

• Leukemia stem cells
• signaling
• Bayesian network
• Neoplasia
• Signal transduction

Acknowledgements

Michael Franklin, a scientific writing editor provided editorial assistance. We extend our appreciation to the core resources at the University of Minnesota that were instrumental in this project: The Hematological Malignancies Tissue Bank which is supported by the National Cancer Institute (5P30CA077598-18); Minnesota Masonic Charities, and the Killebrew-Thompson Memorial Fund through the Cancer Experimental Therapeutics Initiative (CETI); the Flow Cytometry Resource and other services of the Masonic Cancer Center (which is supported by NIH P30 CA77598); the Mass Cytometry Shared Resource at the University of Minnesota which is supported by the Office of the Vice President for Research at the University of Minnesota, and the Minnesota Supercomputing Institute.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

Z. Sachs was supported by: 1. The American Cancer Society, Frederick A. DeLuca Foundation, Mentored Research Scholar Grant (MRSG-16-195-01-DDC). 2. The Lois and Richard King Assistant Professorship in Medicine at the University of Minnesota, the Clinical and Translational Science Institute at the University of Minnesota KL2 Career Development Award and K to R01 Award (NIH/NCATS ULI RR033183 & KL2 RR0333182). 3. The American Cancer Society Institutional Research Grant at the University of Minnesota (124166-IRG-58-001-55-IRG12) 4. The Masonic Cancer Center at the University of Minnesota Translational Working Group Award and Genetic Mechanisms of Cancer Award. 5. The University of Minnesota Department of Medicine Women’s Early Research Career Award. 6. The division of Hematology, Oncology, and Transplantation, Department of Medicine; and the University of Minnesota Foundation donors. 7. The Mugee Research Award for Pediatric Cancer, Masonic Cancer Center, University of Minnesota A. Kay was supported by University of Minnesota Life Sciences Summer Undergraduate Research Programs and NCI R25CA200508. The University of Minnesota’s CTSI is supported through the National Institutes of Health (NIH) Clinical and Translational Science Award (CTSA) program. D. Chang was supported by The University of Minnesota’s CTSI is supported through the National Institutes of Health (NIH) Clinical and Translational Science Award (CTSA) program, grants TL1TR002493 and UL1TR002494. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.