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Blood, Vol. 110, Issue 3, 1013-1021, August 1, 2007
The JAK2 617V>F mutation triggers erythropoietin hypersensitivity and terminal erythroid amplification in primary cells from patients with polycythemia vera
Blood Dupont et al.
110: 1013
Supplemental materials for Dupont et al, Vol 110, Issue 3, 1013-1021
Files in this Data Supplement:
- Figure S1 Comparison and correlation of the JAK2 617V>F/total JAK2 ratio(PDF, 153 KB) -
(A) Quantification of the JAK2 617V>F/total JAK2 ratio in ET and PV granulocyte and bone marrow mononuclear cell DNA. The JAK2 617V>F/total JAK2 DNA ratio in bone marrow mononuclear cells (BM) or granulocytes (GR) from ET and PV patients was determined using real-time PCR techniques. Each lozenge represents one sample. Open bars indicate the median value. (B) Correlation of the JAK2 617V>F/total JAK2 ratio with biological features at the time of diagnosis in PV (black dots) and ET (red dots) patients.
- Figure S2. Epo hypersensitivity of erythroid progenitors from JAK2 617V>F mutated ET and PV patients (PDF, 15 KB) -
CD34+CD38+ cells from JAK2 617V>F positive and normal bone marrow were seeded for a standard methylcellulose assay with various concentrations of Epo. Erythroid colonies were counted, individually picked, and then genotyped by allele-specific real-time PCR. The relative percentage of colonies was defined as the ratio of the absolute number of colonies at a given Epo concentration to the total number of colonies at the maximum concentration or plateau dose (0.5 IU/mL). White bars: wild-type JAK2; grey bars: heterozygous JAK2 617V>F; black bars: homozygous JAK2 617V>F. (A) Genotypic patterns of three representative ET patients. (B) Genotypic patterns of three representative PV patients.
- Figure S3. Selective advantage for mutated cells during erythropoiesis and granulopoiesis (PDF, 18.9 KB) -
The JAK2 617V>F/total JAK2 ratio in immature and mature hematopoietic cells from five ET and seven PV patients was determined by real-time quantitative allele-specific PCR. From the left to the right, each histogram represents the JAK2 617V>F/total JAK2 value in immature cells (CD34+CD38− cells and B/NK/M clones), committed progenitor cells (CD34+CD38+ cells, BFU-E, and CFU-G) and mature cells (erythroblasts, granulocytes) from each patient. Gray histograms: ET patients; hatched histograms: PV patient with a heterozygous profile; black histograms: PV patients with a homozygous profile. The P values were determined using the paired Student t-test.
- Figure S4. A multiple hit model for erythropoiesis in JAK2 617V>F ET and PV (PDF, 117 KB) -
Although alternative events leading to Epo hypersensitivity may exist, the effects of the JAK2 617V>F mutation target terminal erythroid differentiation. An additional molecular defect may enhance the amplification of early progenitors leading to clonality. Various possibilities depending on when these events occur in the hematopoietic stem cell are shown.
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