Extending JAK2V617F and MPLW515 Mutation Analysis to Single Hematopoietic Colonies and B- and T-lymphocytes

¹ Divisions of Hematology, Mayo Clinic, Rochester, MN, USA
² Laboratory Medicine & Pathology, Mayo Clinic, Rochester, MN, USA
³ Dana-Farber Cancer Institute and Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA, and the Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA

^* To whom correspondence should be addressed. E-mail: tefferi.ayalew{at}mayo.edu.

	Abstract

JAK2V617F and MPLW515L/K are myeloproliferative disorder (MPD)-associated mutations. We genotyped 552 individual hematopoietic colonies obtained by CD34+ cell culture, from 16 affected patients (13 JAK2V617F and 3 MPLW515L/K), to determine i) the proportion of colonies harboring a particular mutation in the presence or absence of cytokines, ii) the lineage-distribution of endogenous colonies for each mutation, and iii) the differences (if any) in the pattern of mutation amongst the various MPDs, as established by genotyping of individual colonies. Genotyping analysis revealed cohabitation of mutation-negative and mutation-positive endogenous colonies in polycythemia vera as well as other MPDs. Culture of progenitor cells harboring MPLW515L/K yielded virtually no endogenous erythroid colonies, in contrast to JAK2V617F-harboring progenitor cells. The mutation pattern (i.e. relative distribution of homozygous, heterozygous, or wild-type colonies) was not a distinguishing feature amongst the MPDs, and MPLW515 mutations were detected in B and/or T lymphocytes in all 3 patients tested. These observations suggest that clonal myelopoiesis antedates acquisition of JAK2V617F or MPLW515L/K mutations, and that the latter is acquired in a lympho-myeloid progenitor cell.

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