29 Zahedan Journal of Research in Medical Sciences Journal homepage: www.zjrms.ir Synergistic Effect of Sodium Butyrate and Thalidomide in the Induction of Fetal Hemoglobin Expression in Erythroid Progenitors Derived from Cord Blood CD133 + Cells Ali Dehghanifard, 1 Saeid Kaviani,* 1 Mehrdad Noruzinia, 2 Masoud Soleimani, 1 Saeid Abroun, 1 Abbas Hajifathali, 3 Ali Akbar Pourfathollah, 4 Yousef Mortazavi, 5 Mohammad Ahmadvand, 6 Maryam Mahmoodinia-Meymand, 7 Zeinab Kaviani, 8 Majid Farshdousti-Hagh 9 , Najmaldin Saki 10 1. Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2. Department of Medical Genetics, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran 3. Department of Hematology and Oncology, School of Medical Sciences, Shahid Beheshti University, Tehran, Iran 4. Department of Immunology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran 5. Department of Pathology, Zanjan University of Medical Sciences, Zanjan, Iran 6. Department of Laboratory Sciences, Zahedan University of Medical Sciences, Zahedan, Iran 7. Department of Biology, Stem cell Research Center, Sarem Hospital, Tehran, Iran 8. Department of Internal Medicine, Alborz University of Medical Sciences, Karaj, Iran 9. Laboratory of Hematology and Blood Banking, Tabriz University of Medical Sciences, Tabriz, Iran 10. Thalassemia and Hemoglobinopathy Research Center, Jundi Shapur University of Medical Sciences, Ahvaz, Iran Article information Abstract Article history: Received: 26 Jun 2011 Accepted: 29 Aug 2011 Available online: 19 Dec 2011 Background: The use of drugs with the ability to induce production of fetal hemoglobin as a novel therapeutic approach in treating β-Hemoglobinopathies is considered. γ-globin gene expression inducer drugs including sodium butyrate and thalidomide can reduce additional α-globin chains accumulation in erythroid precursors. Materials and Methods: In this experimental study, MACS kit was used to isolate CD133+ cells of umbilical cord blood. Further, the effect of two drugs of thalidomide and sodium butyrate were separately and combined studied on the induction of quantitative expression of β-globin and γ-globin genes in erythroid precursor cells derived from CD133+ stem cells in-vitro. For this purpose, the technique SYBR green Real-time PCR was used. Results: Flow cytometry results showed that approximately 95% of purified cells were CD133+. Real-time PCR results also showed the increased levels of γ-globin mRNA in the cell groups treated with thalidomide, sodium butyrate and combination of drugs as 2.6 and 1.2 and 3.5 times respectively, and for β-globin gene, it is respectively 1.4 and 1.3 and 1.6 times compared with the control group (p<0.05). Conclusion: The study results showed that the mentioned drug combination can act as a pharmaceutical composition affecting the induction of fetal hemoglobin expression in erythroid precursor cells derived from CD133 + cells. Copyright © 2012 Zahedan University of Medical Sciences. All rights reserved. Keywords: CD133+ cells γ-globin gene Sodium butyrate Thalidomide *Corresponding author at: Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran. E-mail: kavianis@modares.ac.ir Introduction -hemoglobinopathies are genetic disorders of β- globin chain that can cause significant mortality in different parts of the world. Beta-thalassemia and sickle cell disease (SCD) are two common cases of this group of disorders that occur following specific mutations in β-globin gene. SCD is created following point mutations in the sixth codon of β-globin gene that causes substitution of glutamic acid for valine [1]. β-thalassemia also resulted from various types of mutations in β-globin gene and result in reduction or lack of production of β- globin chain [1, 2]. Novel therapeutic solutions based on changes in the epigenetic pattern of genes, which increases gene expression of γ-globin and thus, increase of fetal hemoglobin levels (HbF) are highly regarded. It has been seen in patients with SCD and phenotype HPFH (hereditary persistence of fetal hemoglobin), that high levels of HbF can reduce the disease complications [1-5]. γ-globin gene expression inducer drugs, including hydroxyurea, [6] histone deacetylase (HDAC) inhibitors drugs, such as sodium butyrate [7] and azacitidine [8] and decitabine [9] as well as immunomodulator drugs such as pomalidomide and lenalidomide [10] and thalidomide [11] can reduce the accumulation of additional α-globin chains in erythroid precursors and recovery of non- equilibrium status of alpha chains to non- alpha chains ratio [12]. Various studies have been conducted on the use of growth factors and drugs inducing HbF expression in erythroid precursors derived from hematopoietic stem cells [13-16]. It is seen that CD133+ stem cells are more immature than CD34+ stem cells and has a higher colongenicity potential [17]. While increase of low amounts of HbF in SCD patients can be very effective, increase of significant amounts of HbF in β-thalassemia is required to improve disease symptoms [18]. Therefore, the uses of drugs with high potential in the HbF induction β