single-cell stat5 signal transduction profiling in normal and leukemic stem and progenitor cell populations reveals highly distinct cytokine responses单细胞stat5信号转导分析在正常和白血病干细胞和祖细胞数量显示高度不同的细胞因子的反应.pdfVIP

Single-Cell STAT5 Signal Transduction Profiling in Normal and Leukemic Stem and Progenitor Cell Populations Reveals Highly Distinct Cytokine Responses 1,2 1 1 1 1 Lina Han , Albertus T. J. Wierenga , Marjan Rozenveld-Geugien , Kim van de Lande , Edo Vellenga , Jan Jacob Schuringa1* 1 Department of Hematology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands, 2 Department of Hematology, The First Clinical College of Harbin Medical University, Harbin, China Abstract Background: Signal Transducer and Activator of Transcription 5 (STAT5) plays critical roles in normal and leukemic hematopoiesis. However, the manner in which STAT5 responds to early-acting and lineage-restricted cytokines, particularly in leukemic stem/progenitor cells, is largely unknown. Methodology/Principal Findings: We optimized a multiparametric flow cytometry protocol to analyze STAT5 phosphorylation upon cytokine stimulation in stem and progenitor cell compartments at a single-cell level. In normal cord blood (CB) cells, STAT5 phosphorylation was efficiently induced by TPO, IL-3 and GM-CSF within CD34+CD382 hematopoietic stem cells (HSCs). EPO- and SCF-induced STAT5 phosphorylation was largely restricted to the megakaryocyte-erythroid progenitor (MEP) compartment, while G-CSF as well IL-3 and GM-CSF were most efficient in inducing STAT5 phosphorylation in the myeloid progenitor compartments. Strikingly, mobilized adult peripheral blood (PB) CD34+ cells responded much less efficiently to cytokine-induced STAT5 activation, with the exception of TPO. In leukemi