MicroRNA expression profiling during human cord blood-derived CD34 cell erythropoiesis.
OBJECTIVE
MicroRNA (miRNA) expression profiling was performed on ex vivo differentiating erythroid cultures derived from human umbilical cord blood (UCB) CD34 cells and K562 cells to identify miRNAs involved in erythropoiesis.


METHODS
Both cell types were subjected to growth factor cocktails stimulating erythroid differentiation and were harvested for small RNA extraction at regular intervals. miRNAs with at least a 1.5-fold expression increase or decrease compared to unstimulated (day 0) cells were identified by array hybridization. Validity of the expression array was confirmed by quantitative real-time polymerase chain reaction on randomly selected miRNAs.


RESULTS
Hierarchical clustering analysis and comparison between stimulated UCB-derived CD34 cells and K562 cells revealed miRNAs that are critical for erythroid development and maturation. Correlation analysis on UCB-derived CD34 cells shows that miR-15b, miR-16, miR-22, and miR-185 have strong positive correlation to the appearance of erythroid surface antigens (CD71, CD36, and CD235a) and hemoglobin synthesis, while miR-28 has an inverse relationship to the expression of these markers. Signature miRNAs associated with common myeloid/erythroid progenitor commitment (e.g., miR-181 family, miR-221, miR-154), early erythroid commitment (e.g., miR-32, miR-136, miR-137), and maturation (miR-22, miR-28, miR-185) were also identified by temporal correlation analysis. These miRNAs are predicted to target genes involved in cell development and differentiation.


CONCLUSIONS
Probable signature miRNAs for erythropoiesis are identified. Further experimentations are needed to define the roles of these miRNAs in regulating erythroid commitment.
Choong
Meng Ling
ML
Division of Biomedical Sciences, Singapore Immunology Network, Johns Hopkins in Singapore, 61 Biopolis Drive, Singapore 138673. choong_meng_ling@immunol.a-star.edu.sg
Yang
Henry He
HH
McNiece
Ian
I
D018952:Antigens, CD34 / Q000276:immunology*; D004920:Erythropoiesis; D005312:Fetal Blood / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D020014:K562 Cells; D035683:MicroRNAs / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018952:Antigens, CD34; D035683:MicroRNAs