2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism
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| date | 2023-10-25 |
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| id | 15760 |
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| internalnotes | Sila masukkan subject wajib Dissertations, Academic. Terima kasih... |
| originalfilename | 15760_09b6a5936015c7c.pdf |
| person | Hanan Kamel Mohammed Saad |
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| spelling | 15760 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=15760 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection3 General Document Malaysia Library Staff (Top Management) Library Staff (Management) Library Staff (Support) Terengganu Faculty of Health Sciences English application/pdf 1.5 Server storage Scanned document Universiti Sultan Zainal Abidin UniSZA Private Access UNIVERSITI SULTAN ZAINAL ABIDIN SAMBox 2.3.4; modified using iTextSharp™ 5.5.10 ©2000-2016 iText Group NV (AGPL-version) Copyright©PWB2025 446 15760_09b6a5936015c7c.pdf 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism 2023-10-25 Hemoglobin E-beta thalassemia—Genetics Iron homeostasis is regulated by hepcidin, a hepatic hormone that controls dietary iron absorption and iron recycling. Hepcidin deficiency contributes to iron overload in β Thalassaemia patients. A comprehensive molecular test has not been extensively studied in targeting specific biomarkers for thalassaemia patients. The present study aimed to investigate the dephosphorylation effect of SMAD and JAK/STAT signalling pathways and gene mutation on iron metabolism in HbE/β-Thalassaemia patients and their parents (β-Thalassaemia trait and HbE trait) mother/father. This study is a comparative cross-sectional study conducted with 260 blood samples collected from 65 HbE/β-Thalassaemia patients and 65 parents who attended the Paediatric Unit, Hospital Sultanah Nur Zahirah (HSNZ), and 130 Universiti Sultan Zainal Abidin (UniSZA) students as controls. Complete blood count, iron profile, and serum hepcidin were evaluated. Molecular detection of 21 types of β-Thalassaemia mutations using PCR ARMS technique was performed. Six samples from each group were selected based on the severity, the expression of target genes was evaluated by quantitative reverse transcription PCR (RT-qPCR) analysis. Protein and phosphorylation levels of target proteins were measured using Jess Western analysis. One-way ANOVA, Kruskal Wallis, and Students t-test were conducted for statistical analysis using SPSS software (version 20; IBM Corp), and p<0.05 considered a significant difference. There was a substantial decrease in the number of RBCs, Hb concentration, and RBC indices with significantly decreased serum hepcidin levels and increased serum ferritin in patients and parents (p<0.001) compared to the controls. The gene expression analysis showed a dramatically downregulation of HEPC, sTfR2, HBB, HFE, HJV, VEGFA, BMP2 and BMP4 in patients and parents (p=0.001) compared with controls. However, compared with controls, there was a significant upregulation of FPN1, sTfR1, NGAL, TGF-β1 and FGF23 in patients and parents (p=0.001). The results also revealed 130 β-Thalassaemia mutations among patients and parents. The two most common mutations were detected among the patient’s group, codon 26 (G>A) with Cd 41/42 (-TTCT) and codon 26 (G>A) with IVS 1-5 (G>C). It was found that the most common mutations included codon 26, Cd41/42, and IVS1-5 were detected among parents. The study also identified five novel mutations among patients; Cd 26 (G>A) with (β0)-Thalassaemia Filipino, IVS 1-2 (T>C) with Cd 26 (G>A), and Cd 41/42 (-TTCT), IVS1-5 (G.C) with Cd 26 (G>A). Additionally, there was a significant decreased in protein levels and phosphorylation of STAT5 and SMAD5 in patients and parents (p=0.001) compared with controls. However, STAT3 and SMAD1 were upregulated in patients and parents compared to controls (p=0.001). The findings of present study showed that some mutations were associated with severe anaemia and iron overload in HbE/β Thalassaemia patients. The results also suggested a significant inhibition of STAT5 and SMAD5 signalling pathways in HbE/β-Thalassaemia patients and their parents which was associated with dramatically down-regulated HEPC resulting in iron overload in HbE/β-Thalassaemia patients and parents. Therefore, activation of both signalling pathways could be a future therapeutic target for the management of iron overload burden in β-Thalassaemia patients. Hanan Kamel Mohammed Saad Dissertations, Academic Sila masukkan subject wajib Dissertations, Academic. Terima kasih... HBB Gene Mutation Hemoglobin E/B-Thalassemia Genetics Thalassemia Molecular Profiling Thesis |
| spellingShingle | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| state | Terengganu |
| subject | Hemoglobin E-beta thalassemia—Genetics Dissertations, Academic |
| summary | Iron homeostasis is regulated by hepcidin, a hepatic hormone that controls dietary iron absorption and iron recycling. Hepcidin deficiency contributes to iron overload in β Thalassaemia patients. A comprehensive molecular test has not been extensively studied in targeting specific biomarkers for thalassaemia patients. The present study aimed to investigate the dephosphorylation effect of SMAD and JAK/STAT signalling pathways and gene mutation on iron metabolism in HbE/β-Thalassaemia patients and their parents (β-Thalassaemia trait and HbE trait) mother/father. This study is a comparative cross-sectional study conducted with 260 blood samples collected from 65 HbE/β-Thalassaemia patients and 65 parents who attended the Paediatric Unit, Hospital Sultanah Nur Zahirah (HSNZ), and 130 Universiti Sultan Zainal Abidin (UniSZA) students as controls. Complete blood count, iron profile, and serum hepcidin were evaluated. Molecular detection of 21 types of β-Thalassaemia mutations using PCR ARMS technique was performed. Six samples from each group were selected based on the severity, the expression of target genes was evaluated by quantitative reverse transcription PCR (RT-qPCR) analysis. Protein and phosphorylation levels of target proteins were measured using Jess Western analysis. One-way ANOVA, Kruskal Wallis, and Students t-test were conducted for statistical analysis using SPSS software (version 20; IBM Corp), and p<0.05 considered a significant difference. There was a substantial decrease in the number of RBCs, Hb concentration, and RBC indices with significantly decreased serum hepcidin levels and increased serum ferritin in patients and parents (p<0.001) compared to the controls. The gene expression analysis showed a dramatically downregulation of HEPC, sTfR2, HBB, HFE, HJV, VEGFA, BMP2 and BMP4 in patients and parents (p=0.001) compared with controls. However, compared with controls, there was a significant upregulation of FPN1, sTfR1, NGAL, TGF-β1 and FGF23 in patients and parents (p=0.001). The results also revealed 130 β-Thalassaemia mutations among patients and parents. The two most common mutations were detected among the patient’s group, codon 26 (G>A) with Cd 41/42 (-TTCT) and codon 26 (G>A) with IVS 1-5 (G>C). It was found that the most common mutations included codon 26, Cd41/42, and IVS1-5 were detected among parents. The study also identified five novel mutations among patients; Cd 26 (G>A) with (β0)-Thalassaemia Filipino, IVS 1-2 (T>C) with Cd 26 (G>A), and Cd 41/42 (-TTCT), IVS1-5 (G.C) with Cd 26 (G>A). Additionally, there was a significant decreased in protein levels and phosphorylation of STAT5 and SMAD5 in patients and parents (p=0.001) compared with controls. However, STAT3 and SMAD1 were upregulated in patients and parents compared to controls (p=0.001). The findings of present study showed that some mutations were associated with severe anaemia and iron overload in HbE/β Thalassaemia patients. The results also suggested a significant inhibition of STAT5 and SMAD5 signalling pathways in HbE/β-Thalassaemia patients and their parents which was associated with dramatically down-regulated HEPC resulting in iron overload in HbE/β-Thalassaemia patients and parents. Therefore, activation of both signalling pathways could be a future therapeutic target for the management of iron overload burden in β-Thalassaemia patients. |
| title | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| title_full | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| title_fullStr | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| title_full_unstemmed | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| title_short | 2023_Molecular Characterisation of HBB Gene Mutation In Haemoglobin E/Β-Thalassaemia Patients and The Effects of The Disease on Genes Expression Related to Iron Metabolism |
| title_sort | 2023_molecular characterisation of hbb gene mutation in haemoglobin e/β-thalassaemia patients and the effects of the disease on genes expression related to iron metabolism |