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Mitochondrial Changes in β0-Thalassemia/Hb E Disease.

Khungwanmaythawee K, Sornjai W, Paemanee A, Jaratsittisin J, Fucharoen S, Svasti S, Lithanatudom P, Roytrakul S, Smith DR - PLoS ONE (2016)

Bottom Line: While a number of factors are known that moderate presentation, these factors do not account for the full spectrum of presentation.Differences in relative mitochondria number and gene expression were seen primarily in day 10 cells.The evidence presented here suggest that there are inherent differences in these cells as early as the erythroid progenitor cell stage, and that maximum deficit is seen coincident with high levels of globin gene expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya campus, 25/25 Phuttamontol Sai 4, Salaya, Nakorn Pathom, 73170, Thailand.

ABSTRACT
The compound β°-thalassemia/Hb E hemoglobinopathy is characterized by an unusually large range of presentation from essentially asymptomatic to a severe transfusion dependent state. While a number of factors are known that moderate presentation, these factors do not account for the full spectrum of presentation. Mitochondria are subcellular organelles that are pivotal in a number of cellular processes including oxidative phosphorylation and apoptosis. A mitochondrial protein enriched proteome was determined and validated from erythroblasts from normal controls and β°-thalassemia/Hb E patients of different severities. Mitochondria were evaluated through the use of mitotracker staining, analysis of relative mitochondrial genome number and evaluation of mitochondrial gene expression in addition to assay of overall cellular redox status through the use of alamarBlue assays. Fifty differentially regulated mitochondrial proteins were identified. Mitotracker staining revealed significant differences in staining between normal control erythroblasts and those from β°-thalassemia/Hb E patients. Differences in relative mitochondria number and gene expression were seen primarily in day 10 cells. Significant differences were seen in redox status as evaluated by alamarBlue staining in newly isolated CD34+ cells. Mitochondria mediate oxidative phosphorylation and apoptosis, both of which are known to be dysregulated in differentiating erythrocytes from β°-thalassemia/Hb E patients. The evidence presented here suggest that there are inherent differences in these cells as early as the erythroid progenitor cell stage, and that maximum deficit is seen coincident with high levels of globin gene expression.

No MeSH data available.


Related in: MedlinePlus

Mitochondria in erythroid precursor cells: activity.Day 7, 10 and 14 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were examined for expression of (a) ATP6, (b) ATP8 or (c) CYTB by real time PCR on days 7, 10 and 14 of culture. (d) A total of 1 x104 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were used in alamarBlue assays. Error bars show ± S.E.M. * p ≤ 0.05, ** p ≤ 0.01, ***p ≤ 0.001.
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pone.0153831.g004: Mitochondria in erythroid precursor cells: activity.Day 7, 10 and 14 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were examined for expression of (a) ATP6, (b) ATP8 or (c) CYTB by real time PCR on days 7, 10 and 14 of culture. (d) A total of 1 x104 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were used in alamarBlue assays. Error bars show ± S.E.M. * p ≤ 0.05, ** p ≤ 0.01, ***p ≤ 0.001.

Mentions: Studies have suggested that mitochondria can be heterogeneous with respect to their activity. We therefore used quantitative PCR to determine relative expression levels of three mitochondrially encoded genes, namely ATP synthase F0 subunits 6 and 8 (ATP6 and ATP8) and mitochondrial cytochrome b (CYTB). Results (Fig 4) showed significant differences during differentiation for all three genes investigated. In particular expression of ATP6 and CTY were reduced in erythroblasts from severe β°-thalassemia patients as compared to normal controls on day 10 of culture. Interestingly both ATP6 and ATP8 were increased in expression in both mild and severe β°-thalassemia as compared to normal controls on day 14 of culture.


Mitochondrial Changes in β0-Thalassemia/Hb E Disease.

Khungwanmaythawee K, Sornjai W, Paemanee A, Jaratsittisin J, Fucharoen S, Svasti S, Lithanatudom P, Roytrakul S, Smith DR - PLoS ONE (2016)

Mitochondria in erythroid precursor cells: activity.Day 7, 10 and 14 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were examined for expression of (a) ATP6, (b) ATP8 or (c) CYTB by real time PCR on days 7, 10 and 14 of culture. (d) A total of 1 x104 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were used in alamarBlue assays. Error bars show ± S.E.M. * p ≤ 0.05, ** p ≤ 0.01, ***p ≤ 0.001.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4836671&req=5

pone.0153831.g004: Mitochondria in erythroid precursor cells: activity.Day 7, 10 and 14 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were examined for expression of (a) ATP6, (b) ATP8 or (c) CYTB by real time PCR on days 7, 10 and 14 of culture. (d) A total of 1 x104 erythroid precursor cells from normal controls (white bars) and from mild (grey bars) and severe (black bars) β°-thalassemia/Hb E patients were used in alamarBlue assays. Error bars show ± S.E.M. * p ≤ 0.05, ** p ≤ 0.01, ***p ≤ 0.001.
Mentions: Studies have suggested that mitochondria can be heterogeneous with respect to their activity. We therefore used quantitative PCR to determine relative expression levels of three mitochondrially encoded genes, namely ATP synthase F0 subunits 6 and 8 (ATP6 and ATP8) and mitochondrial cytochrome b (CYTB). Results (Fig 4) showed significant differences during differentiation for all three genes investigated. In particular expression of ATP6 and CTY were reduced in erythroblasts from severe β°-thalassemia patients as compared to normal controls on day 10 of culture. Interestingly both ATP6 and ATP8 were increased in expression in both mild and severe β°-thalassemia as compared to normal controls on day 14 of culture.

Bottom Line: While a number of factors are known that moderate presentation, these factors do not account for the full spectrum of presentation.Differences in relative mitochondria number and gene expression were seen primarily in day 10 cells.The evidence presented here suggest that there are inherent differences in these cells as early as the erythroid progenitor cell stage, and that maximum deficit is seen coincident with high levels of globin gene expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya campus, 25/25 Phuttamontol Sai 4, Salaya, Nakorn Pathom, 73170, Thailand.

ABSTRACT
The compound β°-thalassemia/Hb E hemoglobinopathy is characterized by an unusually large range of presentation from essentially asymptomatic to a severe transfusion dependent state. While a number of factors are known that moderate presentation, these factors do not account for the full spectrum of presentation. Mitochondria are subcellular organelles that are pivotal in a number of cellular processes including oxidative phosphorylation and apoptosis. A mitochondrial protein enriched proteome was determined and validated from erythroblasts from normal controls and β°-thalassemia/Hb E patients of different severities. Mitochondria were evaluated through the use of mitotracker staining, analysis of relative mitochondrial genome number and evaluation of mitochondrial gene expression in addition to assay of overall cellular redox status through the use of alamarBlue assays. Fifty differentially regulated mitochondrial proteins were identified. Mitotracker staining revealed significant differences in staining between normal control erythroblasts and those from β°-thalassemia/Hb E patients. Differences in relative mitochondria number and gene expression were seen primarily in day 10 cells. Significant differences were seen in redox status as evaluated by alamarBlue staining in newly isolated CD34+ cells. Mitochondria mediate oxidative phosphorylation and apoptosis, both of which are known to be dysregulated in differentiating erythrocytes from β°-thalassemia/Hb E patients. The evidence presented here suggest that there are inherent differences in these cells as early as the erythroid progenitor cell stage, and that maximum deficit is seen coincident with high levels of globin gene expression.

No MeSH data available.


Related in: MedlinePlus