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Papers In Press, published online ahead of print December 1, 2004
J. Biol. Chem, 10.1074/jbc.M408085200
Submitted on July 16, 2004
Revised on November 26, 2004
Accepted on December 1, 2004

Concentrative uptake of cyclic ADP-ribose generated by BST-1+ stroma stimulates proliferation of human hematopoietic progenitors

Marina Podestà, Federica Benvenuto, Anna Pitto, Osvaldo Figari, Andrea Bacigalupo, Santina Bruzzone, Lucrezia Guida, Luisa Franco, Laura Paleari, Nicoletta Bodrato, Cesare Usai, Antonio De Flora, and Elena Zocchi

Dept. Experimental Medicine, University of Genova, Genova 16132

Corresponding Author: ezocchi{at}unige.it

Cyclic ADP-ribose is an intracellular calcium mobilizer generated from NAD+ by several ADP-ribosyl cyclases including ectocellular CD38 and BST-1. cADPR, both exogenously added and paracrinally-produced by a CD38+ feeder layer, has recently been demonstrated to stimulate the in vitro proliferation of human hemopoietic progenitors (HP) and also the in vivo expansion of hemopoietic stem cells. The low density of BST-1 expression on bone marrow (BM) stromal cells and the low specific activity of the enzyme, which is significantly weaker compared to CD38, made it unclear whether cADPR generation by a BST-1+ stroma could stimulate HP proliferation in the BM microenvironment. We developed and characterized two BST-1+ stromal cell lines, expressing an ectocellular cyclase activity similar to that of BST-1+ human mesenchymal stem cells, the precursors of BM stromal cells. Long term co-culture of cord blood-derived HP over these BST-1+ feeders determined their expansion, evaluated as increased colony output. Influx of paracrinally generated cADPR into clonogenic HP was found to be mediated by a concentrative, nitrobenzylthioinosine (NBMPR)- and dipyridamole-inhibitable nucleoside transporter, this providing a possible explanation to the effectiveness of the hormone-like concentrations of the cyclic nucleotide measured in the medium conditioned by BST-1+ feeders. These results suggest that the BST-1-catalyzed generation of extracellular cADPR, followed by the concentrative uptake of the cyclic nucleotide by HP, may be physiologically relevant processes in normal hemopoiesis.


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