In vitro expansion of erythroid progenitors from polycythemia vera patients leads to decrease in JAK2 V617F allele.
OBJECTIVE
A G>T transversion in a tyrosine kinase JAK2 (V617F) was reported in over 80% of patients with polycythemia vera (PV). Current evidence suggests that JAK2(V617F) somatic mutation is involved in the pathogenesis of PV, as it confers erythropoietin-independent proliferation to erythroid progenitor cells. However, several unanswered questions regarding the essential role of JAK2(V617F) arose as 1) it is not a dominant mutation, 2) it is not PV-specific as it is found in several myeloproliferative disorders, and 3) some ( approximately 20%) PV patients lack the JAK2(V617F) mutation. We investigated the relative frequency of JAK2(V617F) in in vitro-expanded PV progenitors.


METHODS
In vitro expansion of erythroid progenitors from mononuclear cells was optimized. Frequency of JAK2(V617F) allele was measured by using allele-specific real-time polymerase chain reaction. Clonality was performed using established procedure.


RESULTS
In vitro expansion of PV erythroid progenitors and differentiated dendritic cells resulted in a decrease of the frequency of JAK2(V617F) allele compared with granulocytes or CD235(+) erythroid progenitors. Clonality analysis demonstrated that although granulocytes of these PV patients were clonal, expanded erythroid cells were polyclonal. However, in vitro-expanded PV erythroid progenitors still had approximately a twofold increased proliferative capacity in comparison with erythroid progenitors from healthy individuals. Erythropoietin favors the cells without JAK2(V617F) allele. Dendritic cells in one out of three patients remained clonal.


CONCLUSIONS
JAK2(V617F) mutation does not provide a proliferative/survival advantage to the PV clone during in vitro expansion. These data suggest that the JAK2(V617F) mutation plays an important role in the biology of PV, yet it may not be the PV-initiating event.
Gaikwad
Amos
A
Department of Pediatric-Hematology Oncology, Baylor College of Medicine, Houston, TX, USA.
Nussenzveig
Roberto
R
Liu
Enli
E
Gottshalk
Stephen
S
Chang
KoTung
K
Prchal
Josef T
JT
D000483:Alleles; D001483:Base Sequence; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D003470:Culture Media; D017931:DNA Primers; D015672:Erythroid Precursor Cells / Q000473:pathology*; D004921:Erythropoietin / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D011087:Polycythemia Vera / Q000235:genetics / Q000473:pathology*; D016133:Polymerase Chain Reaction
D003470:Culture Media; D017931:DNA Primers; D004921:Erythropoietin; D053614:Janus Kinase 2