Maintaining the self-renewal and differentiation potential of human CD34+ hematopoietic cells using a single genetic element.
Hematopoiesis is a complex process involving hematopoietic stem cell (HSC) self-renewal and lineage commitment decisions that must continue throughout life. Establishing a reproducible technique that allows for the long-term ex vivo expansion of human HSCs and maintains self-renewal and multipotential differentiation will allow us to better understand these processes, and we report the ability of the leukemia-associated AML1-ETO fusion protein to establish such a system. AML1-ETO-transduced human CD34+ hematopoietic cells routinely proliferate in liquid culture for more than 7 months, remain cytokine dependent for survival and proliferation, and demonstrate self-renewal of immature cells that retain both lymphoid and myeloid potential in vitro. These cells continue to express the CD34 cell surface marker and have ongoing telomerase activity with maintenance of telomere ends, however they do not cause leukemia in nonobese diabetic-severe combined immunodeficiency (NOD/SCID) mice. Identification of the signaling pathways that are modulated by AML1-ETO and lead to the self-renewal of immature human progenitor cells may assist in identifying compounds that can efficiently expand human stem and progenitor cells ex vivo.
Mulloy
James C
JC
Cincinnati Children's Hospital Medical Center, 3333 Burnet Ave, Mail Location 7013, Cincinnati, OH 45229, USA. james.mulloy@cchmc.org
Cammenga
Jorg
J
Berguido
Francisco J
FJ
Wu
Kaida
K
Zhou
Ping
P
Comenzo
Raymond L
RL
Jhanwar
Suresh
S
Moore
Malcolm A S
MA
Nimer
Stephen D
SD
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured / Q000637:transplantation; D003114:Colony-Forming Units Assay; D050676:Core Binding Factor Alpha 2 Subunit; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D015514:Oncogene Proteins, Fusion / Q000502:physiology*; D000075142:RUNX1 Translocation Partner 1 Protein; D011993:Recombinant Fusion Proteins / Q000502:physiology; D015203:Reproducibility of Results; D015398:Signal Transduction; D019098:Telomerase / Q000032:analysis; D014157:Transcription Factors / Q000502:physiology*; D014161:Transduction, Genetic
C107844:AML1-ETO fusion protein, human; D018952:Antigens, CD34; D050676:Core Binding Factor Alpha 2 Subunit; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D019098:Telomerase