Gene Expression Analysis Of The Brazilian Type Of Hereditary Persistence Of Fetal Hemoglobin: Identification Of Genes That Could Be Related To γ-globin Activation

Increased γ-globin production and consequent fetal hemoglobin (Hb F, α2γ2) formation is an important modulator of the clinical and hematological features of hemolytic anemias, such as sickle cell disease and β-thalassemia (β-thal). Hb F genes are genetically regulated, but despite numerous studies, the molecular basis of hemoglobin (Hb) switching is not completely understood. Hereditary persistence of fetal Hb (HPFH) is a consequence of impaired switching in adult life, which results in the continued expression of the γ-globin gene. This study was undertaken to identify genes that could be involved in Hb switching and/or maintenance of elevated Hb F levels. Two libraries were constructed using reticulocytes from normal donors and from Brazilian HPFH subjects. Results suggest that the maintenance of Hb F levels could be associated with some gene/protein expression modifications, such as low expression of KLF1, a transcription factor known to contribute to the regulation and modulation of Hb switching, decreased expression of MIER1, known for the recruitment of chromatin remodeling enzymes, and decreased expression of HOOK3. These data suggest new genes that may play a role in globin gene regulation, γ-globin gene expression and augmentation of Hb F levels, and may represent newly-defined cellular pathways for the control of Hb switching in erythroid cells. © Informa Healthcare USA, Inc.376516535Gazouli, M., Katsantoni, E., Kosteas, T., Anagnou, N.P., Persistent fetal-globin expression in adult transgenic mice following deletion of two silencer elements located 3' to the human A-globin gene (2009) Mol Med., 15 (11-12), pp. 415-424Weatherall, D.J., Phenotype-genotype relationships in monogenic disease: Lessons from the thalassaemias (2001) Nat Rev Genet, 2 (4), pp. 245-255Kim, A., Dean, A., Chromatin loop formation in the-globin locus and its role in globin gene transcription (2012) Mol Cells, 34 (1), pp. 1-5Wilber, A., Nienhuis, A.W., Persons, D.A., Transcriptional regulation of fetal to adult hemoglobin switching: New therapeutic opportunities (2011) Blood, 117 (15), pp. 3945-3953Steinberg, M.H., Genetic etiologies for phenotypic diversity in sickle cell anemia (2009) Sci World J., 9, pp. 46-67Ngo, D.A., Aygun, B., Akinsheye, I., Fetal haemoglobin levels and haematological characteristics of compound heterozygotes for Haemoglobin S and deletional hereditary persistence of fetal haemoglobin (2012) Br J Haematol, 156 (2), pp. 259-264Forget, B.G., Molecular basis of hereditary persistence of fetal hemoglobin (1998) Ann NY Acad Sci., 850, pp. 38-44Costa, F.F., Zago, M.A., Cheng, G., Nechtman, J.F., Stoming, T.A., Huisman, T.H.J., The Brazilian type of nondeletional A-fetal hemoglobin has a C!G substitution at nucleotide-195 of the A-globin gene (1990) Blood, 76 (9), pp. 1896-1897Schreiber, R., Goncalves, M.S., Junqueira, M.L., Saad, S.T., Krieger, J.E., Costa, F.F., The A-195 (C!G) mutation in hereditary persistence of fetal hemoglobin is not associated with activation of a reporter gene in vitro (2001) Braz J Med Biol Res., 34 (4), pp. 489-492Takahashi, T., Schreiber, R., Krieger, J.E., Saad, S.T., Costa, F.F., Analysis of the mechanism of action of the Brazilian type (A-195 C!G) of hereditary persistence of fetal hemoglobin (2003) Eur J Haematol, 71 (6), pp. 418-424Da Cunha, A.F., Brugnerotto, A.F., Corat, M.A., High levels of human-globin are expressed in adult mice carrying a transgene of the Brazilian type of hereditary persistence of fetal hemoglobin (A-195) (2009) Hemoglobin, 33 (6), pp. 439-447Roversi, F.M., Cunha, A.F., Lanaro, C., The-195C!G substitution in Brazilian hereditary persistence of fetal hemoglobin decreases NF-E1/YY1 binding and increases PAXI binding to the A globin promoter region (2010) Proceedings of the 52nd Annual American Society of Hematology Meeting, 116 (21), p. 857. , Orlando, FL, USA, December 4-7 2010. BloodDacie, J.V., Lewis, S.M., (1984) Practical Haematology, , Edinburgh: Churchill LivingstoneGoossens, M., Kan, Y.Y., DNA analysis in the diagnosis of hemoglobin disorders (1981) Methods Enzymol, 76, pp. 805-817Hillmann, A., Dunne, E., Kenny, D., XDNA amplification by SMART-PCR and suppression subtractive hybridization (SSH)-PCR (2009) Methods Mol Biol., 496, pp. 223-243Vandesompele, J., De Preter, K., Pattyn, F., Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes (2002) Genome Biol., 3 (7). , RESEARCH0034Ghosh, S., Zang, S., Mitra, P.S., Ghimbovschi, S., Hoffman, E.P., Dutta, S.K., Global gene expression and ingenuity biological functions analysis on PCBs 153 and 138 induced human PBMC in vitro reveals differential mode(s) of action in developing toxicities (2011) Environ Int., 37 (5), pp. 838-857Ewing, B., Green, P., Base-calling of automated sequencer traces using phred. II. Error probabilities (1998) Genome Res., 8 (3), pp. 186-194Sankaran, V.G., Xu, J., Byron, R., A functional element necessary for fetal hemoglobin silencing (2011) N Engl J Med., 365 (9), pp. 807-814Rosok, O., Sioud, M., Discovery of differentially expressed genes: Technical considerations (2007) Methods Mol Biol., 360, pp. 115-129Diatchenko, L., Lau, Y.F., Campbell, A.P., Suppression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries (1996) Proc Natl Acad Sci USA., 93 (12), pp. 6025-6030De Andrade, T.G., Peterson, K.R., Cunha, A.F., Identification of novel candidate genes for globin regulation in erythroid cells containing large deletions of the human-globin gene cluster (2006) Blood Cells Mol Dis., 37 (2), pp. 82-90Bonafoux, B., Lejeune, M., Piquemal, D., Analysis of remnant reticulocyte mRNA reveals new genes and antisense transcripts expressed in the human erythroid lineage (2004) Haematologica, 89 (12), pp. 1434-1438Goh, S.H., Ytbhanunv, L., Anewly discovered human-globin gene (2005) Blood, 106 (4), pp. 1466-1472Silver, N., Best, S., Jiang, J., Thein, S.L., Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR (2006) BMC Mol Biol., 7 (1), pp. 33-42Goh, S.H., Josleyn, M., Lee, Y.T., The human reticulocytes transcriptome (2007) Physiol Genomic., 30 (2), pp. 172-178Wojda, U., Noel, P., Miller, J.L., Fetal and adult hemoglobin production during adult erythropoiesis: Coordinate expression correlates with cell proliferation (2002) Blood, 99 (8), pp. 3005-3013Van Den Berg, M.C., Burgering, B.M., Integrating opposing signals toward Forkhead box O (2011) Antioxid Redox Signal, 14 (4), pp. 607-621Monsalve, M., Olmos, Y., The complex biology of FOXO (2011) Curr Drug Targets, 12 (9), pp. 1322-1350Ding, Z., Gillespie, L.L., Mercer, F.C., Paterno, G.D., The SANT domain of human MI-ER1 interacts with Sp1 to interfere with GC box recognition and repress transcription from its own promoter (2004) J Biol Chem., 279 (27), pp. 28009-28016Blackmore, T.M., Mercer, C.F., Paterno, G.D., Gillespie, L.L., The transcriptional cofactor MIER1-negatively regulates histone acetyltransferase activity of the CREB-binding protein (2008) BMC Res Notes, 1 (1), pp. 68-73Kaiser, F., Kaufmann, S.H., Zerrahn, J., IIGP, a member of the IFN inducible and microbial defense mediating 47 kDa GTPase family, interacts with the microtubule binding protein hook3 (2004) J Cell Sci., 117 (PART. 9), pp. 1747-1756Chatterjee, M., Agrawal, S., Agarwal, S.S., Differential effect of IFN-and IFN-on phosphorylation of p65 and p50 (rel) in the K562 cell line: Implications for altered interaction with RXR (1996) Cytokine, 8 (5), pp. 357-364Bauer, D.E., Kamran, S.C., Orkin, S.H., Reawakening fetal hemoglobin: Prospects for new therapies for the-globin disorders (2012) Blood, 120 (15), pp. 2945-2953Roversi, F.M., Cunea, A.F., Brugnerotto, A.F., Lanaro, C., Albuquerque, D.M., Costa, F.F., Gene expression profile analysis in hereditary persistence of fetal hemoglobin (Brazilian type): Identification of target genes that could be related to hemoglobin switching (2011) Proceedings of the 16th Congress of the European Haematology Association, 96 (SUPPL. 2), pp. 169-170. , London, UK, June 9-12 2011. HaematologicaJackson, D.A., McDowell, J.C., Dean, A., Globin locus control region HS2 and HS3 interact structurally and functionally (2003) Nucleic Acids Res., 31 (4), pp. 1180-1190Borg, J., Papadopoulos, P., Georgitsi, M., Haploinsufficiency for the erythroid transcription factor KLF1 causes hereditary persistence of fetal hemoglobin (2010) Nat Genet, 42 (9), pp. 801-805Satta, S., Perseu, L., MacCioni, L., Giagu, N., Galanello, R., Delayed fetal hemoglobin switching in subjects with KLF1 gene mutation (2012) Blood Cells Mol Dis., 48 (1), pp. 22-24Zhou, D., Liu, K., Sun, C.W., Pawlik, K.M., Townes, T.M., KLF1 regulates BCL11A expression and-to-globin gene switching (2010) Nat Genet, 42 (9), pp. 742-74

General movements detect early signs of hemiplegia in term infants with neonatal cerebral infarction

Background. Studies have reported that infants with hemiplegia of congenital origin may have a period between birth and up to 12 months when clinical signs of hemiplegia are not evident. The aim of this study was to establish whether the assessment of general movements (GMs) may help in the earlier detection of signs of hemiplegia. Subjects and Methods. Eleven infants with cerebral infarction on brain MRI, and eleven normal controls were enrolled in the study. Quality of GMs was assessed from videotapes between 3 and 6 weeks and between 9 and 16 weeks. Neurological outcome was evaluated at least at two years. Results. Seven of the 11 infants had an assessment performed between 3 and 6 weeks: abnormal GMs were observed in all the infants who developed hemiplegia, but one child had abnormal GMs and a normal outcome. All 11 infants had a scorable assessment between 9 and 16 weeks. In all a specific type of GMs, fidgety movements (FMs), were predictive of neurological outcome. The presence of early asymmetries at both 3 - 6 and 9 - 16 weeks was also significantly associated with later signs of hemiplegia. Conclusions. The assessment of GMs after the neonatal period appears to be very useful in the early identification of hemiplegia in infants with cerebral infarction. Whilst the prediction of hemiplegia should be possible from early neonatal MRI brain scans, this facility is not always available. Observation of GMs is a bedside clinical approach that allows confirmation of early prediction from MRI, early rehabilitation if needed and reassurance that neurological outcome will be good where that is appropriat