Pumilio1 (PUM1) Expression, Sickle Cell Anemia, β-thalassemia Intermedia

Официальное название

Detection of RNA Binding Protein Pumilio1 (PUM1) Expression in Patients With Sickle Cell Anemia and β-thalassemia Intermedia

Обзор

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.

Подробное описание

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.

Вмешательства

• Диагностический тест RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR
  Quantatative Real -Time PCR for quantification of PUM1; * The type of the recruited samples: Ethylenediamine tetra-acetic acid (EDTA) peripheral blood samples. * The method of total RNA extraction: TRIzol and TRIzol LS. * The purity and concentration of the RNA were measured using a Nano Drop 2000 instrument. * cDNA was amplified with primers using the GoScript Reverse Transcription system. Western Blotting Assay of PUM1 protein levels

Первичные конечные точки

• Expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia. [Срок оценки: 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]

Критерии участия

Критерии включения

• Patients with sickle cell anemia and β-thalassemia intermedia.
• Patients are of both sexes (males and females) at any age.

Критерии исключения

• Patients with any other type of haemolytic anaemias.
• Patients on Hydroxyurea therapy.

Критерии приведены из реестра в оригинале (на английском). Окончательную оценку соответствия проводит исследовательский центр.

Дизайн исследования

Модель наблюдения

Случай-контроль

Центры проведения

Список центров уточняется — проверьте первичный протокол.

Публикации

• Persons DA. Hematopoietic stem cell gene transfer for the treatment of hemoglobin disorders. Hematology Am Soc Hematol Educ Program. 2009:690-7. doi: 10.1182/asheducation-2009.1.690. PMID 20008255
• Sankaran VG, Orkin SH. The switch from fetal to adult hemoglobin. Cold Spring Harb Perspect Med. 2013 Jan 1;3(1):a011643. doi: 10.1101/cshperspect.a011643. PMID 23209159
• Sankaran VG, Nathan DG. Reversing the hemoglobin switch. N Engl J Med. 2010 Dec 2;363(23):2258-60. doi: 10.1056/NEJMcibr1010767. No abstract available. PMID 21121839
• Nuinoon M, Makarasara W, Mushiroda T, Setianingsih I, Wahidiyat PA, Sripichai O, Kumasaka N, Takahashi A, Svasti S, Munkongdee T, Mahasirimongkol S, Peerapittayamongkol C, Viprakasit V, Kamatani N, Winichagoon P, Kubo M, Nakamura Y, Fucharoen S. A genome-wide association identified the common genetic variants influence disease severity in beta0-thalassemia/hemoglobin E. Hum Genet. 2010 Mar;127(3): PMID 20183929
• Stamatoyannopoulos G. Control of globin gene expression during development and erythroid differentiation. Exp Hematol. 2005 Mar;33(3):259-71. doi: 10.1016/j.exphem.2004.11.007. PMID 15730849
• Elagooz R, Dhara AR, Gott RM, Adams SE, White RA, Ghosh A, Ganguly S, Man Y, Owusu-Ansah A, Mian OY, Gurkan UA, Komar AA, Ramamoorthy M, Gnanapragasam MN. PUM1 mediates the posttranscriptional regulation of human fetal hemoglobin. Blood Adv. 2022 Dec 13;6(23):6016-6022. doi: 10.1182/bloodadvances.2021006730. PMID 35667093
• Goldstrohm AC, Hall TMT, McKenney KM. Post-transcriptional Regulatory Functions of Mammalian Pumilio Proteins. Trends Genet. 2018 Dec;34(12):972-990. doi: 10.1016/j.tig.2018.09.006. Epub 2018 Oct 10. PMID 30316580

Идентификаторы

NCT: NCT05883254 · PUM1 Sickle cell Thalassemia

Первоисточники (государственные реестры)

• ClinicalTrials.gov (NIH, США)

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