The long-term objective of the research is to develop reagents that activate human gamma globin gene transcription in the adult for treatment of beta-thalassemia and sickle cell anemia. It is known that expression of the fetal gamma globin genes greatly ameliorates the effects of these diseases, however it is not known how the human gamma globin genes are normally suppressed (or expressed at very low levels) when the expression of the human gamma globin genes switches to that of the delta and beta globin genes. It could be the result of an absence of transcription factors required for gamma globin gene expression or the presence of active suppressors of gamma globin expression or both.
The aim of this application is therefore twofold: the characterization of the stage specific factors that are required for gamma globin gene expression and/or the stage specific factors that suppress gamma globin gene expression. Two independent in vivo approaches will be used. One uses chromatin precipitation to directly purify and sequence the proteins bound to the gamma globin gene promoters in the fetal stage and adult stage. The other is a functional assay to obtain single chain antibodies that switch on the gamma globin genes in the adult stage.
|Pourfarzad, Farzin; von Lindern, Marieke; Azarkeivan, Azita et al. (2013) Hydroxyurea responsiveness in ?-thalassemic patients is determined by the stress response adaptation of erythroid progenitors and their differentiation propensity. Haematologica 98:696-704|
|Pourfarzad, Farzin; Aghajanirefah, Ali; de Boer, Ernie et al. (2013) Locus-specific proteomics by TChP: targeted chromatin purification. Cell Rep 4:589-600|
|Papadopoulos, Petros; Gutiérrez, Laura; van der Linden, Reinier et al. (2012) A dual reporter mouse model of the human ?-globin locus: applications and limitations. PLoS One 7:e51272|
|Giardine, Belinda; Borg, Joseph; Higgs, Douglas R et al. (2011) Systematic documentation and analysis of human genetic variation in hemoglobinopathies using the microattribution approach. Nat Genet 43:295-301|
|Borg, Joseph; Papadopoulos, Petros; Georgitsi, Marianthi et al. (2010) Haploinsufficiency for the erythroid transcription factor KLF1 causes hereditary persistence of fetal hemoglobin. Nat Genet 42:801-5|
|van Dijk, Thamar Bryn; Gillemans, Nynke; Pourfarzad, Farzin et al. (2010) Fetal globin expression is regulated by Friend of Prmt1. Blood 116:4349-52|
|Patrinos, George P; Grosveld, Frank G (2008) Pharmacogenomics and therapeutics of hemoglobinopathies. Hemoglobin 32:229-36|
|Papachatzopoulou, Adamantia; Kaimakis, Polynikis; Pourfarzad, Farzin et al. (2007) Increased gamma-globin gene expression in beta-thalassemia intermedia patients correlates with a mutation in 3'HS1. Am J Hematol 82:1005-9|
|Rodriguez, Patrick; Bonte, Edgar; Krijgsveld, Jeroen et al. (2005) GATA-1 forms distinct activating and repressive complexes in erythroid cells. EMBO J 24:2354-66|