Transforming growth factor-beta1 causes transcriptional activation of CD34 and preserves haematopoietic stem/progenitor cell activity.
Stem/progenitor cells endowed with in vitro and in vivo haematopoietic activity express the surface protein CD34. Transforming growth factor beta1 (TGF-beta1) is one of the soluble molecules that regulate cell cycle and differentiation of haematopoietic cells, but has pleiotropic activities depending on the state of responsiveness of the target cells. It has previously been shown that TGF-beta1 maintains human CD34+ haematopoietic progenitors in an undifferentiated state, independently of any cell cycle effect. Here, we have shown that TGF-beta1 upregulates the human CD34, an effect that was evident in primary stem/progenitor cells (CD34+lin-) both at the transcriptional and protein levels, and was not associated with any relevant effect on cell growth. The presence of TGF-beta1 influenced differentiation, maintaining primary CD34+/Lin- in an undifferentiated state. This effect was associated with Smad activation and with a dramatic decrease in p38 phosphorylation. Moreover, blocking p38 phosphorylation by the SB202190 inhibitor increased CD34 RNA levels but did not enhance CD34 protein expression in CD34+/Lin- cells, suggesting that modulation of multiple signalling pathways is necessary to reproduce TGF-beta1 effects. These data establish the role that TGF-beta1 has in the modulation of the CD34 stem/progenitor protein and stem/progenitor functions, providing important clues for understanding haematopoietic development and a potential tool for the modulation of human haematopoiesis.
Istituto di Ematologia, Dipartimento per la Salute della Donna e della Vita Nascente, Universitá Cattolica del Sacro Cuore, Largo A. Gemelli 8, 00168 Rome, Italy. luca@pierelli.it
Luca
L
Pierelli
Maria
M
Marone
Giuseppina
G
Bonanno
Sergio
S
Rutella
Daniela
D
de Ritis
Salvatore
S
Mancuso
Giuseppe
G
Leone
Giovanni
G
Scambia
D018952:Antigens, CD34 / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D007093:Imidazoles / Q000037:antagonists & inhibitors; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010766:Phosphorylation; D011725:Pyridines / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015533:Transcriptional Activation; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1; D048051:p38 Mitogen-Activated Protein Kinases
D018952:Antigens, CD34; D007093:Imidazoles; D011725:Pyridines; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole