TY - JOUR
T1 - Comparative, genome-scale transcriptional analysis of CHRF-288-11 and primary human megakaryocytic cell cultures provides novel insights into lineage-specific differentiation
AU - Fuhrken, Peter G.
AU - Chen, Chi
AU - Miller, William M.
AU - Papoutsakis, Eleftherios T.
N1 - Funding Information:
This work was supported by a National Institutes of Health grant (HL48276) and the Robert H. Lurie Comprehensive Cancer Center. P.F. was supported by a National Science Foundation Graduate Research Fellowship. We acknowledge the use of instruments in the Keck Biophysics Facility, the Biological Imaging Facility, and the Center for Genetic Medicine at Northwestern University. We gratefully acknowledge Genentech for the donation of TPO. Finally, we thank Lisa Giammona and Carlos Paredes for helpful discussions and assistance.
PY - 2007/3
Y1 - 2007/3
N2 - Objectives: Little is known about the transcriptional events underlying megakaryocytic (Mk) differentiation. We sought to identify genes and pathways previously unassociated with megakaryopoiesis and to evaluate the CHRF-288-11 (CHRF) megakaryoblastic cell line as a model system for investigating megakaryopoiesis. Methods: Using DNA microarrays, Q-RT-PCR, and protein-level assays, we compared the dynamic gene expression pattern of phorbol ester-induced differentiation of CHRF cells to cytokine-induced Mk differentiation of human mobilized peripheral blood CD34+ cells. Results: Transcriptional patterns of well-known Mk genes were similar between the two systems. CHRF cells constitutively express some early Mk genes including GATA-1. Expression patterns of apoptosis-related genes suggested that increased p53 activity is involved in Mk apoptosis, and this was confirmed by p53-DNA-binding activity data and flow-cytometric analysis of the p53 target gene BBC3. Certain Rho and G-protein-coupled-receptor signaling pathway components were upregulated, including genes not previously associated with Mk cells. Ontological analysis revealed upregulation of defense-response genes, including both known and candidate platelet-derived contributors to inflammation. Upregulation of interferon-responsive genes occurred in the cell line, but not in the primary cells, likely due to a known genetic mutation in the JAK2/STAT5 signaling pathway. Conclusions: This analysis of megakaryopoiesis, which integrates dynamic gene expression data with protein abundance and activity assays, has identified a number of genes and pathways that may help govern megakaryopoiesis. Furthermore, the transcriptional data support the hypothesis that CHRF cells resemble an early Mk phenotype and, with certain limitations, exhibit genuine transcriptional features of Mk differentiation upon treatment with phorbol esters.
AB - Objectives: Little is known about the transcriptional events underlying megakaryocytic (Mk) differentiation. We sought to identify genes and pathways previously unassociated with megakaryopoiesis and to evaluate the CHRF-288-11 (CHRF) megakaryoblastic cell line as a model system for investigating megakaryopoiesis. Methods: Using DNA microarrays, Q-RT-PCR, and protein-level assays, we compared the dynamic gene expression pattern of phorbol ester-induced differentiation of CHRF cells to cytokine-induced Mk differentiation of human mobilized peripheral blood CD34+ cells. Results: Transcriptional patterns of well-known Mk genes were similar between the two systems. CHRF cells constitutively express some early Mk genes including GATA-1. Expression patterns of apoptosis-related genes suggested that increased p53 activity is involved in Mk apoptosis, and this was confirmed by p53-DNA-binding activity data and flow-cytometric analysis of the p53 target gene BBC3. Certain Rho and G-protein-coupled-receptor signaling pathway components were upregulated, including genes not previously associated with Mk cells. Ontological analysis revealed upregulation of defense-response genes, including both known and candidate platelet-derived contributors to inflammation. Upregulation of interferon-responsive genes occurred in the cell line, but not in the primary cells, likely due to a known genetic mutation in the JAK2/STAT5 signaling pathway. Conclusions: This analysis of megakaryopoiesis, which integrates dynamic gene expression data with protein abundance and activity assays, has identified a number of genes and pathways that may help govern megakaryopoiesis. Furthermore, the transcriptional data support the hypothesis that CHRF cells resemble an early Mk phenotype and, with certain limitations, exhibit genuine transcriptional features of Mk differentiation upon treatment with phorbol esters.
UR - http://www.scopus.com/inward/record.url?scp=33847044392&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33847044392&partnerID=8YFLogxK
U2 - 10.1016/j.exphem.2006.10.017
DO - 10.1016/j.exphem.2006.10.017
M3 - Article
C2 - 17309828
AN - SCOPUS:33847044392
SN - 0301-472X
VL - 35
SP - 476-489.e23
JO - Experimental Hematology
JF - Experimental Hematology
IS - 3
ER -
