primitive myelofibrosis Evidence for MPL W515L/K mutations in hematopoietic stem cells in

ABSTRACT The MPL (W515L and W515K) mutations have been detected in granulocytes of patients suffering from certain types of Primitive Myelofibrosis (PMF). It is still unknown whether this molecular event is also present in lymphoid cells and therefore potentially at the Hematopoietic stem cell (HSC) level. Towards this goal, we conducted MPL genotyping of mature myeloid and lymphoid cells and of lymphoid/myeloid progenitors isolated from PMF patients carrying the W515 mutations. We detected both MPL mutations in granulocytes, monocytes and platelets as well as NK cells but not in T cells. B/NK/Myeloid and/or NK/Myeloid CD34 + CD38 - derived clones were found to carry the mutations. Long term reconstitution of MPL W515 CD34 + cells in NOD/SCID mice was successful as long as 12 weeks post-transplantation indicating that MPL W515 mutations were present in HSC. Moreover, the two MPL mutations induced a spontaneous megakaryocytic growth in culture with an overall normal response to TPO. In contrast, erythroid progenitors remained EPO-dependent. These results demonstrate that in PMF, the MPL W515L or K mutation induces a spontaneous MK differentiation and occurs in a multipotent HSC.

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