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This article is part of the supplement: 18th Scientific Symposium of the Austrian Pharmacological Society (APHAR)

Open Access Meeting abstract

Pharmacological stimulation of murine and human hematopoetic stem cells

Christian Bergmayr1, Christian Balasz1, Zarzar Kazemi1, Filza Hussain1, Thomas Bauer2, Herbert Strobl2, Peter Selzer3, Wolfgang Strohmaier4, Michael Freissmuth1 and Eva Zebedin-Brandl1*

Author affiliations

1 Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University of Vienna, 1090 Vienna, Austria

2 Institute of Immunology, Medical University of Vienna, 1090 Vienna, Austria

3 Department of Radiation Oncology, Medical University of Vienna, 1090 Vienna, Austria

4 SciPharm SàRL, 6131 Junglinster, Luxembourg

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Citation and License

BMC Pharmacology and Toxicology 2012, 13(Suppl 1):A67  doi:10.1186/2050-6511-13-S1-A67


The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/2050-6511/13/S1/A67


Published:17 September 2012

© 2012 Bergmayr et al; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Background

Hematopoietic stem cell (HSC) transplantation is a standard procedure in the treatment of hematological disorders and also applicable to support aggressive chemotherapy in cancer. In practice, the clinical outcome is often limited by inefficient bone marrow (BM) engraftment or by low numbers of available stem cells. It would therefore be desirable to enhance engraftment by pharmacological stimulation. HSC require several signals for successful migration into the bone marrow. One of these signals is provided by stimulation of Gαs[1]. Pretreatment with prostaglandin (PG) E2 enhances engraftment via activation of Gαs-coupled EP2 and EP4 receptors [2]. Treprostinil is a stable analogue of prostacyclin/PGI2, which acts via IP, EP2 and EP4 receptors and is approved for treatment of pulmonary hypertension. Here we tested the hypothesis that treprostinil stimulates stem cell engraftment.

Methods

Generation of murine bone marrow-derived HSCs: Undifferentiated HSC (lineage-negative, Lin− cells) were isolated from murine BM, separated by MACS (magnetic-assisted cell sorting) and characterized by fluorescence-activated cell sorting (FACS). BM transplantation: Lin− cells were pretreated in vitro in the absence and presence of 10 µM treprostinil alone or in combination with 30 µM FSK for 1 h at 37°C. The cellular threshold for successful transplantation was determined by titrating the number of HSCs required to allow for survival of lethally irradiated recipient mice. Engraftment and transplantation efficiency was determined by the analysis of white blood cell counts.

Results

Trepostinil triggered a concentration-dependent accumulation of cAMP in murine Lin− cells with an estimated EC50 in the range of 0.3 µM. A treprostini-induced cAMP accumulation was also observed in human HSCs, which were also shown to express the mRNA encoding IP, EP2 and EP4 receptors. Treprostinil enhanced engraftment of HSCs; this conclusion is based on the following observations: (i) mice injected with treprostinil-pretreated Lin− cells had significantly higher levels of circulating white blood cells as compared to those receiving vehicle-treated Lin− cells (p < 0.05, unpaired t-test); (ii) when pretreated and untreated Lin− cells were mixed to compete for BM reconstitution, the pretreatment with treprostinil increased transplantation efficiency 1.5–3-fold; (iii) in vitro pretreatment of HSCs with trepostinil reduced the minimum number of cells required to rescue a mouse; (iv) engraftment of HSCs was further enhanced when treprostinil was also administered after injection of the pretreated HSCs.

Conclusions

Treprostinil is suitable for improving haematopoetic stem cell transplantaion. The observations allow for designing a protocol to test the compound in a clinical trial.

References

  1. Adams GB, Alley IR, Chung UI, Chabner KT, Jeanson NT, Lo Celso C, Marsters ES, Chen M, Weinstein LS, Lin CP, Kronenberg HM, Scadden DT: Haematopoietic stem cells depend on Gαs-mediated signalling to engraft bone marrow.

    Nature 2009, 459:103-107. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  2. Hoggatt J, Singh P, Sampath J, Pelus LM: Prostaglandin E2 enhances hematopoietic stem cell homing, survival, and proliferation.

    Blood 2009, 113:5444-5455. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL