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Restoration of Haemoglobin Level Using Hydrodynamic Gene Therapy with Erythropoietin Does Not Alleviate the Disease Progression in an Anaemic Mouse Model for TGFβ1-Induced Chronic Kidney Disease.

Pedersen L, Wogensen L, Marcussen N, Cecchi CR, Dalsgaard T, Dagnæs-Hansen F - PLoS ONE (2015)

Bottom Line: Epo exerts its haematopoietic effects by stimulating the proliferation and differentiation of erythrocytes with subsequent improved tissue oxygenation.Epo receptors are furthermore expressed in non-haematopoietic tissue and today, Epo is recognised as a cytokine with many pleiotropic effects.Thus, Epo treatment in this model of chronic kidney disease normalises haemoglobin levels but has no effect on kidney fibrosis or function.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, Aarhus, Denmark.

ABSTRACT
Erythropoietin, Epo, is a 30.4 kDa glycoprotein hormone produced primarily by the fetal liver and the adult kidney. Epo exerts its haematopoietic effects by stimulating the proliferation and differentiation of erythrocytes with subsequent improved tissue oxygenation. Epo receptors are furthermore expressed in non-haematopoietic tissue and today, Epo is recognised as a cytokine with many pleiotropic effects. We hypothesize that hydrodynamic gene therapy with Epo can restore haemoglobin levels in anaemic transgenic mice and that this will attenuate the extracellular matrix accumulation in the kidneys. The experiment is conducted by hydrodynamic gene transfer of a plasmid encoding murine Epo in a transgenic mouse model that overexpresses TGF-β1 locally in the kidneys. This model develops anaemia due to chronic kidney disease characterised by thickening of the glomerular basement membrane, deposition of mesangial matrix and mild interstitial fibrosis. A group of age matched wildtype littermates are treated accordingly. After a single hydrodynamic administration of plasmid DNA containing murine EPO gene, sustained high haemoglobin levels are observed in both transgenic and wildtype mice from 7.5 ± 0.6 mmol/L to 9.4 ± 1.2 mmol/L and 10.7 ± 0.3 mmol/L to 15.5 ± 0.5 mmol/L, respectively. We did not observe any effects in the thickness of glomerular or tubular basement membrane, on the expression of different collagen types in the kidneys or in kidney function after prolonged treatment with Epo. Thus, Epo treatment in this model of chronic kidney disease normalises haemoglobin levels but has no effect on kidney fibrosis or function.

No MeSH data available.


Related in: MedlinePlus

Evaluation of plasmid expression site and level.A: Plasmid signal day 8 after injection in Tg Control (n = 8) and Wt Control (n = 5) corresponding to the location of the liver. Scanning Control mice without preceding injection with luciferin are scanned simultaneously (n = 4). B: Regions of interest marks the signal and is measured as total flux (photons/second) (± SD). There is a tendency to higher plasmid expression in Wt Control mice (n = 5) than Tg Control mice (n = 8) (P = 0.13). C: Expression of Epo mRNAfrom the liver at termination of the experiment. Both Tg (n = 11) and Wt (n = 11) mice treated with Epo express significant more plasmid from the liver compared to their Controls (¤,§P < 0.001, ± SD). Significant difference between Tg Epo 1.25 μg/mouse (n = 11) and Wt Epo 1.25 μg/mouse (n = 11) (#P < 0.001, ± SD) are found.
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pone.0128367.g002: Evaluation of plasmid expression site and level.A: Plasmid signal day 8 after injection in Tg Control (n = 8) and Wt Control (n = 5) corresponding to the location of the liver. Scanning Control mice without preceding injection with luciferin are scanned simultaneously (n = 4). B: Regions of interest marks the signal and is measured as total flux (photons/second) (± SD). There is a tendency to higher plasmid expression in Wt Control mice (n = 5) than Tg Control mice (n = 8) (P = 0.13). C: Expression of Epo mRNAfrom the liver at termination of the experiment. Both Tg (n = 11) and Wt (n = 11) mice treated with Epo express significant more plasmid from the liver compared to their Controls (¤,§P < 0.001, ± SD). Significant difference between Tg Epo 1.25 μg/mouse (n = 11) and Wt Epo 1.25 μg/mouse (n = 11) (#P < 0.001, ± SD) are found.

Mentions: To confirm plasmid expression in the liver, we injected a group of Tg and Wt littermates with Control plasmid following the same procedure as described above. After 8 days the mice were injected intraperitoneal with luciferin (Tg n = 8, Wt n = 5). The mice were visualised and light emission was measured. As expected, the light emission was observed in mice treated with Control plasmid corresponding to the location of the liver. No light emission was seen in the Scanning Control without injection of luciferin (Fig 2A and 2B). Wildtype mice had a tendency to express more luciferase than Tg mice, however, this was not statistically significant (P = 0.13).


Restoration of Haemoglobin Level Using Hydrodynamic Gene Therapy with Erythropoietin Does Not Alleviate the Disease Progression in an Anaemic Mouse Model for TGFβ1-Induced Chronic Kidney Disease.

Pedersen L, Wogensen L, Marcussen N, Cecchi CR, Dalsgaard T, Dagnæs-Hansen F - PLoS ONE (2015)

Evaluation of plasmid expression site and level.A: Plasmid signal day 8 after injection in Tg Control (n = 8) and Wt Control (n = 5) corresponding to the location of the liver. Scanning Control mice without preceding injection with luciferin are scanned simultaneously (n = 4). B: Regions of interest marks the signal and is measured as total flux (photons/second) (± SD). There is a tendency to higher plasmid expression in Wt Control mice (n = 5) than Tg Control mice (n = 8) (P = 0.13). C: Expression of Epo mRNAfrom the liver at termination of the experiment. Both Tg (n = 11) and Wt (n = 11) mice treated with Epo express significant more plasmid from the liver compared to their Controls (¤,§P < 0.001, ± SD). Significant difference between Tg Epo 1.25 μg/mouse (n = 11) and Wt Epo 1.25 μg/mouse (n = 11) (#P < 0.001, ± SD) are found.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4457485&req=5

pone.0128367.g002: Evaluation of plasmid expression site and level.A: Plasmid signal day 8 after injection in Tg Control (n = 8) and Wt Control (n = 5) corresponding to the location of the liver. Scanning Control mice without preceding injection with luciferin are scanned simultaneously (n = 4). B: Regions of interest marks the signal and is measured as total flux (photons/second) (± SD). There is a tendency to higher plasmid expression in Wt Control mice (n = 5) than Tg Control mice (n = 8) (P = 0.13). C: Expression of Epo mRNAfrom the liver at termination of the experiment. Both Tg (n = 11) and Wt (n = 11) mice treated with Epo express significant more plasmid from the liver compared to their Controls (¤,§P < 0.001, ± SD). Significant difference between Tg Epo 1.25 μg/mouse (n = 11) and Wt Epo 1.25 μg/mouse (n = 11) (#P < 0.001, ± SD) are found.
Mentions: To confirm plasmid expression in the liver, we injected a group of Tg and Wt littermates with Control plasmid following the same procedure as described above. After 8 days the mice were injected intraperitoneal with luciferin (Tg n = 8, Wt n = 5). The mice were visualised and light emission was measured. As expected, the light emission was observed in mice treated with Control plasmid corresponding to the location of the liver. No light emission was seen in the Scanning Control without injection of luciferin (Fig 2A and 2B). Wildtype mice had a tendency to express more luciferase than Tg mice, however, this was not statistically significant (P = 0.13).

Bottom Line: Epo exerts its haematopoietic effects by stimulating the proliferation and differentiation of erythrocytes with subsequent improved tissue oxygenation.Epo receptors are furthermore expressed in non-haematopoietic tissue and today, Epo is recognised as a cytokine with many pleiotropic effects.Thus, Epo treatment in this model of chronic kidney disease normalises haemoglobin levels but has no effect on kidney fibrosis or function.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, Aarhus, Denmark.

ABSTRACT
Erythropoietin, Epo, is a 30.4 kDa glycoprotein hormone produced primarily by the fetal liver and the adult kidney. Epo exerts its haematopoietic effects by stimulating the proliferation and differentiation of erythrocytes with subsequent improved tissue oxygenation. Epo receptors are furthermore expressed in non-haematopoietic tissue and today, Epo is recognised as a cytokine with many pleiotropic effects. We hypothesize that hydrodynamic gene therapy with Epo can restore haemoglobin levels in anaemic transgenic mice and that this will attenuate the extracellular matrix accumulation in the kidneys. The experiment is conducted by hydrodynamic gene transfer of a plasmid encoding murine Epo in a transgenic mouse model that overexpresses TGF-β1 locally in the kidneys. This model develops anaemia due to chronic kidney disease characterised by thickening of the glomerular basement membrane, deposition of mesangial matrix and mild interstitial fibrosis. A group of age matched wildtype littermates are treated accordingly. After a single hydrodynamic administration of plasmid DNA containing murine EPO gene, sustained high haemoglobin levels are observed in both transgenic and wildtype mice from 7.5 ± 0.6 mmol/L to 9.4 ± 1.2 mmol/L and 10.7 ± 0.3 mmol/L to 15.5 ± 0.5 mmol/L, respectively. We did not observe any effects in the thickness of glomerular or tubular basement membrane, on the expression of different collagen types in the kidneys or in kidney function after prolonged treatment with Epo. Thus, Epo treatment in this model of chronic kidney disease normalises haemoglobin levels but has no effect on kidney fibrosis or function.

No MeSH data available.


Related in: MedlinePlus