TY - JOUR
T1 - MIR-23A microRNA cluster inhibits B-cell development
AU - Kong, Kimi Y.
AU - Owens, Kristin S.
AU - Rogers, Jason H.
AU - Mullenix, Jason
AU - Velu, Chinavenmeni S.
AU - Grimes, H. Leighton
AU - Dahl, Richard
N1 - Funding Information:
We would like to thank Brandy Comyford and Shannon Fitzpatrick for their technical assistance, and Ed Bedrick for statistics consultation. The MSCV-EGFP vector was generously provided by Rodney DeKoter (University of Western Ontario, London, ON, Canada). G1E-ER4 cells were a generous gift from Mitch Weiss (University of Pennsylvania, Philadelphia, PA, USA). 32Dcl3 cells were provided by Allan Friedman (Johns Hopkins, Baltimore, MD, USA). This work was supported by an American Cancer Society Research Scholar Grant (Atlanta, GA, USA) ( RSG-06-170-01-LIB , R.D.), an American Society of Hematology Junior Faculty Scholar Grant (Washington, DC, USA) (R.D.), and a grant from the dedicated health research funds of the University of New Mexico School of Medicine (Albuquerque, NM, USA) (R.D.). J.M. was partially supported by an Institutional Research Training Award from the American Society of Hematology .
PY - 2010/8
Y1 - 2010/8
N2 - Objective: The transcription factor PU.1 (encoded by Sfpi1) promotes myeloid differentiation, but it is unclear what downstream genes are involved. Micro RNAs (miRNAs) are a class of small RNAs that regulate many cellular pathways, including proliferation, survival, and differentiation. The objective of this study was to identify miRNAs downstream of PU.1 that regulate hematopoietic development. Materials and Methods: miRNAs that change expression in a PU.1-inducible cell line were identified with microarrays. The promoter for an miRNA cluster upregulated by PU.1 induction was analyzed for PU.1 binding by electrophoretic mobility shift and chromatin immunoprecipitation assays. Retroviral transduction of hematopoietic progenitors was performed to evaluate the effect of miRNA expression on hematopoietic development in vitro and in vivo. Results: We identified an miRNA cluster whose pri-transcript is regulated by PU.1. The pri-miRNA encodes three mature miRNAs: miR-23a, miR-27a, and miR-24-2. Each miRNA is more abundant in myeloid cells compared to lymphoid cells. When hematopoietic progenitors expressing the 23a cluster miRNAs were cultured in B-cell-promoting conditions, we observed a dramatic decrease in B lymphopoiesis and an increase in myelopoiesis compared to control cultures. In vivo, hematopoietic progenitors expressing the miR-23a cluster generate reduced numbers of B cells compared to control cells. Conclusions: The miR-23a cluster is a downstream target of PU.1 involved in antagonizing lymphoid cell fate acquisition. Although miRNAs have been identified downstream of PU.1 in mediating development of monocytes and granulocytes, the 23a cluster is the first downstream miRNA target implicated in regulating development of myeloid vs lymphoid cells.
AB - Objective: The transcription factor PU.1 (encoded by Sfpi1) promotes myeloid differentiation, but it is unclear what downstream genes are involved. Micro RNAs (miRNAs) are a class of small RNAs that regulate many cellular pathways, including proliferation, survival, and differentiation. The objective of this study was to identify miRNAs downstream of PU.1 that regulate hematopoietic development. Materials and Methods: miRNAs that change expression in a PU.1-inducible cell line were identified with microarrays. The promoter for an miRNA cluster upregulated by PU.1 induction was analyzed for PU.1 binding by electrophoretic mobility shift and chromatin immunoprecipitation assays. Retroviral transduction of hematopoietic progenitors was performed to evaluate the effect of miRNA expression on hematopoietic development in vitro and in vivo. Results: We identified an miRNA cluster whose pri-transcript is regulated by PU.1. The pri-miRNA encodes three mature miRNAs: miR-23a, miR-27a, and miR-24-2. Each miRNA is more abundant in myeloid cells compared to lymphoid cells. When hematopoietic progenitors expressing the 23a cluster miRNAs were cultured in B-cell-promoting conditions, we observed a dramatic decrease in B lymphopoiesis and an increase in myelopoiesis compared to control cultures. In vivo, hematopoietic progenitors expressing the miR-23a cluster generate reduced numbers of B cells compared to control cells. Conclusions: The miR-23a cluster is a downstream target of PU.1 involved in antagonizing lymphoid cell fate acquisition. Although miRNAs have been identified downstream of PU.1 in mediating development of monocytes and granulocytes, the 23a cluster is the first downstream miRNA target implicated in regulating development of myeloid vs lymphoid cells.
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U2 - 10.1016/j.exphem.2010.04.004
DO - 10.1016/j.exphem.2010.04.004
M3 - Article
C2 - 20399246
AN - SCOPUS:77954536744
SN - 0301-472X
VL - 38
SP - 629-640.e1
JO - Experimental Hematology
JF - Experimental Hematology
IS - 8
ER -