Pumilio1 (PUM1) Expression, Sickle Cell Anemia, β-thalassemia Intermedia
- Conditions
- Sickle Cell Disease, Beta Thalassemia Intermedia
- Interventions
- Diagnostic Test: RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR
- Registration Number
- NCT05883254
- Lead Sponsor
- Assiut University
- Brief Summary
1. To study the expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia.
2. To detect PUM1 protein levels in sickle cell anemia and β-thalassemia intermedia patients.
3. To correlate PUM1 gene expression pattern and protein levels with HbF levels in sickle cell anemia and β-thalassemia intermedia patients.
- Detailed Description
The disorders of β-hemoglobin, sickle cell disease (SCD) and β-thalassemia, are major causes of morbidity and mortality worldwide. These diseases are the most common genetic disorders in the world.
SCD is due to a single base-pair point mutation in the β-globin gene resulting in the substitution of valine for glutamic acid in the β-globin chain. The pathophysiology is directly related to polymerization of deoxygenated hemoglobin, leading to a cascade of pathologic events including erythrocyte sickling, vaso-occlusion, and hemolysis.
In β-thalassemia, insufficient production of the β-globin molecule results in an excess of free α-globin chains that can precipitate within erythroid precursors, impairing their maturation and leads to death of these precursors and ineffective production of erythroid cells. As a result, a significant anemia occurs and the consequent expansion of erythroid precursors can lead to secondary problems in bones and other organs.
The hemoglobin molecule is a tetramer composed of two subunits of α-like globin peptides and two subunits of the β-like globin peptides, along with heme moieties. β-globin switching from fetal γ-globin (HBG1 and HBG2) to adult β-globin is a developmental process that occurs in erythrocytes at around the time of birth. Fetal hemoglobin (HbF) induction in adult erythrocytes is an effective therapeutic strategy for SCD and β-thalassemia.
Pumilio1 (PUM1) is a novel target of the erythroid master transcription factor erythroid Krüppel-like factor (EKLF). PUM1 is a member of Pumilio-Fem3-binding factor (PUF) family of sequence specific RNA-binding proteins, acts as a posttranscriptional repressor by binding to the 3' untranslated region (3'-UTR) of messenger RNA (mRNA). It peaks during terminal erythroid differentiation and binds to fetal γ-globin (HBG1) mRNA and impairs its stability and translation. HBG1 has 2 core PUM1 consensus binding sites, but HBG2 and adult globins do not. Knockdown of PUM1 leads to a robust increase in HBF (∼22%) without affecting β-globin levels in human erythroid cells. Moreover, targeting PUM1 does not affect erythropoiesis, which provides a potentially safe and effective therapeutic strategy for SCD and β-thalassemia. Also it was found that elevated HbF levels in the absence of anemia in an individual with a novel heterozygous PUM1 mutation in the RNA-binding domain, which suggests that PUM1 is a critical player during human hemoglobin switching.
Recruitment & Eligibility
- Status
- NOT_YET_RECRUITING
- Sex
- All
- Target Recruitment
- 80
- Patients with sickle cell anemia and β-thalassemia intermedia.
- Patients are of both sexes (males and females) at any age.
- Patients with any other type of haemolytic anaemias.
- Patients on Hydroxyurea therapy.
Study & Design
- Study Type
- OBSERVATIONAL
- Study Design
- Not specified
- Arm && Interventions
Group Intervention Description Control group RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR Normal individuals thalassemia intermedia patients and sickle cell disease patients RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR 1. Inclusion criteria * Patients with sickle cell anemia and β-thalassemia intermedia. * Patients are of both sexes (males and females) at any age. 2. Exclusion criteria: * Patients with any other type of haemolytic anaemias. * Patients on Hydroxyurea therapy.
- Primary Outcome Measures
Name Time Method Expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia. saple taken after diagnosis and before recieve treatment. If patients already on treatment , sample taken at least one month after stoppage of hydroxyurea or blood transfusion Detect PUM1 protein levels in sickle cell anemia and β-thalassemia intermedia patients and to correlate PUM1 gene expression pattern and protein levels with HbF levels in sickle cell anemia and β-thalassemia intermedia patients.
- Secondary Outcome Measures
Name Time Method