Clonal selection and asymmetric distribution of human leukemia in murine xenografts revealed by cellular barcoding

Mirjam E. Belderbos, Taco Koster, Bertien Ausema, Sabrina Jacobs, Sharlaine Sowdagar, Erik Zwart, Eveline de Bont, Gerald de Haan and Leonid V. Bystrykh

Key Points

  • Patient-derived leukemia xenografts are highly polyclonal and show stochastic and clone-size–driven selection.

  • Leukemia clones are asymmetrically distributed in xenografts and preferentially localize to the bone marrow or to extramedullary sites.

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Genetic and phenotypic heterogeneity of human leukemia is thought to drive leukemia progression through a Darwinian process of selection and evolution of increasingly malignant clones. However, the lack of markers that uniquely identify individual leukemia clones precludes high-resolution tracing of their clonal dynamics. Here, we use cellular barcoding to analyze the clonal behavior of patient-derived leukemia-propagating cells (LPCs) in murine xenografts. Using a leukemic cell line and diagnostic bone marrow cells from 6 patients with B-progenitor cell acute lymphoblastic leukemia, we demonstrate that patient-derived xenografts were highly polyclonal, consisting of tens to hundreds of LPC clones. The number of clones was stable within xenografts but strongly reduced upon serial transplantation. In contrast to primary recipients, in which clonal composition was highly diverse, clonal composition in serial xenografts was highly similar between recipients of the same donor and reflected donor clonality, supporting a deterministic, clone-size–based model for clonal selection. Quantitative analysis of clonal abundance in several anatomic sites identified 2 types of anatomic asymmetry. First, clones were asymmetrically distributed between different bones. Second, clonal composition in the skeleton significantly differed from extramedullary sites, showing similar numbers but different clone sizes. Altogether, this study shows that cellular barcoding and xenotransplantation providea useful model to study the behavior of patient-derived LPC clones, which provides insights relevant for experimental studies on cancer stem cells and for clinical protocols for the diagnosis and treatment of leukemia.

  • Submitted December 20, 2016.
  • Accepted March 27, 2017.
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