Investigation of transcription factors involved in HBG1 gene expression regulation employing transgenic beta-YAC mice and CD34+ human cells models with Brazilian type of non-deletional hereditary persistence of fetal hemoglobin

• Grant number: 14/17413-9
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: December 30, 2014
• End date: December 22, 2015
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Fernando Ferreira Costa
• Grantee: Carolina Ayumi Braghini
• Supervisor: Kenneth Richard Peterson
• Host Institution: Centro de Hematologia e Hemoterapia (HEMOCENTRO). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: University of Kansas Medical Center (KUMC), United States
• Associated to the scholarship: 12/07869-0 - Investigation of the role of YY1 and PAX1 transcription factors in the molecular mechanisms involved in the Brazilian type of hereditary persistence of fetal hemoglobin, BP.DR

Abstract

Hereditary persistence of fetal hemoglobin (HPFH) is a condition characterized by elevated synthesis of g-globin in adult definitive erythroid cells, which normally have only very low levels of fetal hemoglobin (HbF), due to a failure in the hemoglobin switch from HbF to adult hemoglobin (HbA). Fetal hemoglobin is a major genetic modifier of the phenotypic heterogeneity in patients with the major ²-globin disorders sickle cell disease and ²-thalassemia. Thus, research has focused on elucidating the pathways involved in the maintenance or activation of HBG1 (gamma globin A) and HBG2 (gamma globin G) gene expression by drug or gene therapy. Many studies have demonstrated the role of stage-specific transcription factors in hemoglobin switching, indicating the potential therapeutic use of these transcription factors to treat hemoglobinopathies. Since there are many HPFH mutations that directly or indirectly affect binding of transcription factors, in this study, we will analyze the proteins which bind the -195 C>G mutation site in the HBG1 gene of Brazilian type nondeletional HPFH, using human ²-globin locus yeast artificial chromosome (²-YAC) transgenic mice and derivative cells, and human CD34+ erythroid progenitor cells. For this study, we will combine stable isotope tagging of erythroid cell nuclear extracts, pull down of interacting proteins with biotin-tagged oligonucleotides, and mass spectrometry (MS) to detect specific HBG1 gene promoter-associated proteins. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BRAGHINI, CAROLINA A.; COSTA, FLAVIA C.; FEDOSYUK, HALYNA; NEADES, RENEE Y.; NOVIKOVA, LESYA V.; PARKER, MATTHEW P.; WINEFIELD, ROBERT D.; PETERSON, KENNETH R.. Generation of non-deletional hereditary persistence of fetal hemoglobin beta-globin locus yeast artificial chromosome transgenic mouse models:-175 Black HPFH and-195 Brazilian HPFH. Experimental Biology and Medicine, v. 241, n. 7, p. 697-705, APR 2016. (14/17413-9)