Sickle Cell Disease, Beta Thalassemia IntermediaNCT05883254
Pumilio1 (PUM1) Expression, Sickle Cell Anemia, β-thalassemia Intermedia
An observational study of Sickle Cell Disease, Beta Thalassemia Intermedia, run by Assiut University.
Trial 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. Read more about this study
- Who is it for?
- Any age
- Conditions the study is for
- Sickle Cell Disease, Beta Thalassemia Intermedia
- What is being tested?
- RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR
- How many people
- 80 participants
- Where
- No sites listed yet
- Study dates
- Aug 2023to Dec 2027
- Phase
- Observational
- Are there results?
- Not yet: the study is under way
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages Any age
- All sexes
- Child, Adult, Older adult
Study population: Patients with sickle cell anemia and β-thalassemia intermedia
Who may be able to join
- Patients with sickle cell anemia and β-thalassemia intermedia.
- Patients are of both sexes (males and females) at any age.
Who cannot join
- Patients with any other type of haemolytic anaemias.
- Patients on Hydroxyurea therapy.
Trial locations and contacts
No sites are listed yet. Contact the study team or a site near you to ask about taking part.
About this study
Official title: Detection of RNA Binding Protein Pumilio1 (PUM1) Expression in Patients With Sickle Cell Anemia and β-thalassemia Intermedia
- To study the expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia.
- To detect PUM1 protein levels in sickle cell anemia and β-thalassemia intermedia patients.
- To correlate PUM1 gene expression pattern and protein levels with HbF levels in sickle cell anemia and β-thalassemia intermedia patients.
Read the 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.
- Being tested
- RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR
- Medical subject headings
- Anemia, Sickle Cell, beta-Thalassemia
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCRDiagnostic test
Also called Western Blotting Assay of PUM1 protein levels
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
| Study group | Role | What they receive |
|---|---|---|
| Control group | Normal individuals Receives: Diagnostic Test: RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR | |
| thalassemia intermedia patients and sickle cell disease patients | 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. Receives: Diagnostic Test: RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR |
What is being measured
The primary outcome is the main question the study is designed to answer.
Expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia.
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.
Measured: 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
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted31 May 2023
- Study startAug 2023 (estimated)
- Primary completionDec 2026 (estimated)
- Study completionDec 2027 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (7)Papers the sponsor has linked to this study.
- Other trials for Sickle Cell Disease, Beta Thalassemia IntermediaStudies of the same condition, open and under way.
- More trials run by Assiut UniversityThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Enrolment
- 80 participants (estimated)
Sponsor and investigators
- Lead sponsor
- Assiut University
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Mervat Awad Mohamed Ibrahim, Assiut University
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT05883254
- Sponsor's protocol ID
- PUM1 Sickle cell Thalassemia
- Status last verified
- May 2023
- First submitted
- 21 May 2023
- Last updated
- 31 May 2023
- Results posted
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
Publications
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.
PubMed 20008255Sankaran 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.
PubMed 23209159Sankaran 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.
PubMed 21121839Nuinoon 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):303-14. doi: 10.1007/s00439-009-0770-2.
PubMed 20183929Stamatoyannopoulos 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.
PubMed 15730849Elagooz 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.
PubMed 35667093Goldstrohm 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.
PubMed 30316580
Study at a glance
From the registry record, last updated by the sponsor on 31 May 2023. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Detection of RNA Binding Protein Pumilio1 (PUM1) Expression in Patients With Sickle Cell Anemia and β-thalassemia Intermedia
- ClinicalTrials.gov ID
- NCT05883254
- Acronym / sponsor ID
- PUM1 Sickle cell Thalassemia
- Study type
- Observational
- Recruitment status
- Not yet recruiting
- Enrolment
- 80 participants (estimated)
- Lead sponsor
- Assiut UniversityUniversity, hospital or other
- Study start
- Aug 2023 (estimated)
- Primary completion
- Dec 2026 (estimated)
- Study completion
- Dec 2027 (estimated)
- First posted
- 31 May 2023 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
- To study the expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia.
- To detect PUM1 protein levels in sickle cell anemia and β-thalassemia intermedia patients.
- 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.
Study design
- Observational model
- Case-control
- Time perspective
- Cross-sectional
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| Control group | – | Diagnostic Test: RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCRNormal individuals |
| thalassemia intermedia patients and sickle cell disease patients | – | Diagnostic Test: RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCR1. 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. |
Who may qualify
- Sex
- All
- Age groups
- Child, Adult, Older adult
- Study population
- Patients with sickle cell anemia and β-thalassemia intermedia
Key inclusion criteria (2 listed)
- Patients with sickle cell anemia and β-thalassemia intermedia.
- Patients are of both sexes (males and females) at any age.
Key exclusion criteria (2 listed)
- Patients with any other type of haemolytic anaemias.
- Patients on Hydroxyurea therapy.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| RNA Binding Protein Pumilio1 (PUM1) Expression by reverse transcriptase quantatative PCRAlso: Western Blotting Assay of PUM1 protein levelsQuantatative 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 | Diagnostic test | Control group, thalassemia intermedia patients and sickle cell disease patients |
Primary outcome
Expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia.
Time frame: 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.
Results and safety
This study is still under way. Results are not available in the source record yet.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 31 May 2023.
- Background: PubMed 20008255Persons 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.
- Background: PubMed 23209159Sankaran 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.
- Background: PubMed 21121839Sankaran 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.
- Background: PubMed 20183929Nuinoon 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):303-14. doi: 10.1007/s00439-009-0770-2.
- Background: PubMed 15730849Stamatoyannopoulos 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.
- Background: PubMed 35667093Elagooz 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.
- Background: PubMed 30316580Goldstrohm 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.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part.
Questions about this trial
What is this clinical trial studying?
An observational study of Sickle Cell Disease, Beta Thalassemia Intermedia, run by Assiut University.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 2 inclusion and 2 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: opening soon, as of the sponsor's last update on 31 May 2023. Status can change between updates, so confirm with the study contact before making plans.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
This study is still under way. Results are not available in the source record yet.
How can a doctor refer a patient or contact the research team?
Contact Mervat Awad Mohamed, Master (01011893938, Mervat_Awad@aun.edu.eg), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Expression pattern of PUM1 gene in patients with sickle cell anemia and β-thalassemia intermedia. (time frame: 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). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT05883254, which also lists 7 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 31 May 2023.