Induction of fetal hemoglobin (HbF) by novel alkylating agents in human erythroid cells; synergistic effects with established HbF-inducing agents

 
see the original item page
in the repository's web site and access all digital files if the item*
share




2008 (EN)
Induction of fetal hemoglobin (HbF) by novel alkylating agents in human erythroid cells; synergistic effects with established HbF-inducing agents

Mourelatos, Dionysios
Mioglou-Kalouptsi, Eleftheria
Chiotoglou, I.
Liakos, P.
Kollia, P.
Samara, M.
Likousi, S.
Papadakis, M.
Chassanidis, C.
Samara, S.
Iakovidou-Kritsi, Zafeiroula

Pharmacological induction of fetal hemoglobin (HbF) is beneficial for some patients with β-thalassemia and also ameliorates the severity of pain episodes in sickle cell anemia, mainly by hydroxyurea (HU). However, refractoriness or poor response of some patients treated with HU triggered research for other drugs. In the present study, we evaluated the effects of novel steroidal alkylating agents EA80, XK4 and CS-211 on HbF induction in CD34+ cell cultures from normal donors. Furthermore, we examined the effects of these agents combined with HU, hemin (HE) and butyric acid (BU) on HbF modulation in adult erythroid cells. CD34+ cells from normal donors cultured in serum-free StemSpan medium were exposed to EPO (4 u/ml) + SCF (100 ng/ml). Different concentrations of EA80 (0.1–1 μM), XK4 (0.1–10 μM) and CS-211 (0.1–10 μM) were added to the cultures at day 5 (proerythroblast stage), then cells were washed and harvested 1–3 days later. The effect of the drugs on cell number was measured by the trypan blue exclusion technique. The number of Hb-containing cells was determined using the benzidine-HCl procedure. EA80, over a wide range of concentrations (0.1–0.8 μM), did not compromise cell survival. Continuous exposure of CD34+ to EA80 had a dose (up to 0.4 μM)- and -time dependent effect on cell number as well as on globin mRNA levels. Treatment of CD34+ cells with EA80 at 0.4 μM for 3 days was followed by a twofold increase in cell number and a 1.5-fold in benzidine-positive cells. Qualitative and quantitative RT-PCR evaluation of globin-mRNA transcripts in CD34+ demonstrated that EA80 (0.4 μÌ) caused a time- and dose-dependent increase in gamma globin mRNA (1–3 days: 1.5 to 2.0-fold). The addition of HU and HE in combination with EA80 in normal CD34+ cell cultures led to a 20–30% increase in cell number by day 9. Furthermore, the combination of EA80 with either HU or HE was accompanied by an increase in γ-mRNA content (1.5- and 2.5-fold, respectively). The addition of BU in combination with EA80 had the predominant effect on erythroid cell proliferation and γ-mRNA levels (fivefold at day 3). The addition of XK4 at any concentrations proved toxic (reduced cell number and γ-globin mRNA transcripts). The addition of the third drug, CS-211, at concentrations between 0.1 and 10 μM had a dose (up to 5 μÌ)- and -time dependent effect on cell number and γ-globin mRNA transcripts [highest effect at day 3, 4.0-fold at 5 μÌ]. The level of both adult globin mRNAs was not affected by any of the tested drugs. Our findings suggest that the beneficial effect of EA80 and CS-211 might be threefold: (i) increasing cell number, (ii) affecting preferentially the rate of transcription of γ-globin mRNA, (iii) acting synergistically with HE, HU and BU, most likely through transcriptional and posttranscriptional mechanisms. These results indicate that novel alkylating agents EA80 and CS-211, either alone or in combination with other HbF-augmenting drugs, might provide a potentially useful treatment for patients with β-hemoglobinopathies with poor or no response to established Hb-F inducing agents

Article / Άρθρο
info:eu-repo/semantics/article

Αριστοτέλειο Πανεπιστήμιο Θεσσαλονίκης (EL)
Aristotle University of Thessaloniki (EN)

2009-07-17T11:41:39Z
2008


Αριστοτέλειο Πανεπιστήμιο Θεσσαλονίκης, Σχολή Επιστημών Υγείας, Τμήμα Ιατρικής

urn:ISSN:10799796
Blood Cells Molecules and Diseases, vol.40 [2008] p.260-261 [Published Version]

This record is part of 'IKEE', the Institutional Repository of Aristotle University of Thessaloniki's Library and Information Centre found at http://ikee.lib.auth.gr. Unless otherwise stated above, the record metadata were created by and belong to Aristotle University of Thessaloniki Library, Greece and are made available to the public under Creative Commons Attribution-ShareAlike 4.0 International license (http://creativecommons.org/licenses/by-sa/4.0). Unless otherwise stated in the record, the content and copyright of files and fulltext documents belong to their respective authors. Out-of-copyright content that was digitized, converted, processed, modified, etc by AUTh Library, is made available to the public under Creative Commons Attribution-ShareAlike 4.0 International license (http://creativecommons.org/licenses/by-sa/4.0). You are kindly requested to make a reference to AUTh Library and the URL of the record containing the resource whenever you make use of this material.
info:eu-repo/semantics/openAccess



*Institutions are responsible for keeping their URLs functional (digital file, item page in repository site)