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 Blood, 1 August 2008, Vol. 112, No. 3, pp. 886-890.
Prepublished online as a Blood First Edition Paper on June 5, 2008; DOI 10.1182/blood-2008-02-138909.

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RED CELLS

Brief Report

Attenuation of EPO-dependent erythroblast formation by death-associated protein kinase-2

Jing Fang1, Madhu Menon1, Diya Zhang2, Bruce Torbett3, Leif Oxburgh1, Mario Tschan3, Estelle Houde1, and Don M. Wojchowski1

1 Stem and Progenitor Cell Biology Program, Molecular Medicine Division, Maine Medical Center Research Institute, Scarborough; 2 Department of Statistics, Northwestern University, Evanston, IL; and 3 Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA

The adult erythron is maintained via dynamic modulation of erythroblast survival potentials. Toward identifying novel regulators of this process, murine splenic erythroblasts at 3 developmental stages were prepared, purified and profiled. Stage-to-stage modulated genes were then functionally categorized, with a focus on apoptotic factors. In parallel with BCL-X and NIX, death-associated protein kinase-2 (DAPK2) was substantially up-modulated during late erythropoiesis. Among hematopoietic lineages, DAPK2 was expressed predominantly in erythroid cells. In a Gata1-IE3.9int-DAPK2 transgenic mouse model, effects on steady-state reticulocyte and red blood cell (RBC) levels were limited. During hemolytic anemia, however, erythropoiesis was markedly deficient. Ex vivo ana-lyses revealed heightened apoptosis due to DAPK2 at a Kit–CD71highTer119– stage, together with a subsequent multifold defect in late-stage Kit–CD71highTer119+ cell formation. In UT7epo cells, siRNA knock-down of DAPK2 enhanced survival due to cytokine withdrawal, and DAPK2's phosphorylation and kinase activity also were erythropoietin (EPO)-modulated. DAPK2 therefore comprises a new candidate attenuator of stress erythropoiesis.


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