1. Myeloid leukemia with transdifferentiation plasticity developing from T‐cell progenitors
Representative FACS plot (left) showing a mixed‐lineage clone with a T‐lymphoid (CD25+/CD11b−Gr1−), myeloid (CD11b+Gr1+/CD25−), and biphenotypic (CD11b+Gr1+/CD25+) fraction derived from a single Myc/Bcl2‐transformed DN2 cell grown on 1:1 OP9:OP9‐Dll1 co‐culture with lineage‐promiscuous cytokines IL‐2, IL‐3, IL‐6, IL‐7, SCF, GM‐CSF, Flt3. Morphology of the indicated flow‐sorted cell fractions was analyzed by Giemsa staining (right). Phenotype frequencies of clones from Myc/Bcl2‐transformed single cells (grown and phenotyped as mentioned in A). Analyzed were 31 clones from LSKs, 107 from GMPs and 28 from DN2 cells. Mice that received 5 × 104 to 5 × 105 T‐lymphoid cells or myeloid cells flow‐sorted from mixed Myc/Bcl2+ DN2‐derived clones showed a cumulative survival of 31 ± 2 and 27 ± 6 days respectively. The summarized data are from three mice injected with the T‐lymphoid and four mice injected with the myeloid fraction of two different clones. Representative FACS plots demonstrating predominant outgrowth of myeloid clones from single Myc/Bcl2‐transformed LSK or GMP cells on OP9:OP9‐Dll1 co‐cultures. (top panels) Serial replating of Myc‐Bcl2+ DN2 cells in methylcellulose generated myeloid clones with stable genomic reassembly of the Dβ1 TCR locus in vitro (gel: left two lanes). Upon transplantation, the same clones induced myeloid leukemia that retained the initial rearrangement in vivo (gel: right two lanes) as assessed by nested PCR. GL = germline. (bottom panels) Methylcellulose‐based replating of Myc‐Bcl2+ GMPs generated myeloid clones that exclusively displayed the TCRβ locus in germline configuration. Cumulative survival of mice that received 106 cells of TCRβ‐rearranged myeloid DN2‐derived clones leading to death 33 ± 4 days after transplantation. Two DN2 clones were transplanted into five recipient mice.
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EMBO J, Volume: 35, Issue: 22, Pages: , First published: 29 August 2016, DOI: ( /embj )