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Transplantation of ATP7B-transduced bone marrow mesenchymal stem cells decreases copper overload in rats.

Chen S, Shao C, Dong T, Chai H, Xiong X, Sun D, Zhang L, Yu Y, Wang P, Cheng F - PLoS ONE (2014)

Bottom Line: IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B.This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all).Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery Ward of General Surgery, The Affiliated Wuhu No. 2 People's Hospital of Wannan Medical College, Wuhu, Anhui Province, China.

ABSTRACT

Background: Recent studies have demonstrated that transplantation of ATP7B-transduced hepatocytes ameliorates disease progression in LEC (Long-Evans Cinnamon) rats, a model of Wilson's disease (WD). However, the inability of transplanted cells to proliferate in a normal liver hampers long-term treatment. In the current study, we investigated whether transplantation of ATP7B-transduced bone marrow mesenchymal stem cells (BM-MSCs) could decrease copper overload in LEC rats.

Materials and methods: The livers of LEC rats were preconditioned with radiation (RT) and/or ischemia-reperfusion (IRP) before portal vein infusion of ATP7B-transduced MSCs (MSCsATP7B). The volumes of MSCsATP7B or saline injected as controls were identical. The expression of ATP7B was analyzed by real-time quantitative polymerase chain reaction (RT-PCR) at 4, 12 and 24 weeks post-transplantation. MSCATP7B repopulation, liver copper concentrations, serum ceruloplasmin levels, and alanine transaminase (ALT) and aspartate transaminase (AST) levels were also analyzed at each time-point post-transplantation.

Results: IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B. This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all). Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).

Conclusions: These results demonstrate that transplantation of MSCsATP7B into IRP-plus-RT preconditioned LEC rats decreased copper overload and was associated with an increase in MSC engraftment and repopulation.

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Related in: MedlinePlus

Transfection efficacy, expression of ATP7B post-transfection and post-transplantation.(A) Transfection efficacy of pRRL.PPT.SF.ATP7B.i2GFPBsd.pre in BM-MSCs. (B) Transfection efficacy of pRRL.PPT.SF.i2GFPpre in BM-MSCs. Transfected cells were observed under both a fluorescence microscope and a regular microscope (×100); (C) Expression of ATP7B in BM-MSCs post-transfection assessed by Western Blot. (D) Expression of ATP7B in each group at each time-point following transplantation in vivo. *p<0.05 compared with the saline group, &p<0.05 compared with IRP and RT group, respectively.
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pone-0111425-g002: Transfection efficacy, expression of ATP7B post-transfection and post-transplantation.(A) Transfection efficacy of pRRL.PPT.SF.ATP7B.i2GFPBsd.pre in BM-MSCs. (B) Transfection efficacy of pRRL.PPT.SF.i2GFPpre in BM-MSCs. Transfected cells were observed under both a fluorescence microscope and a regular microscope (×100); (C) Expression of ATP7B in BM-MSCs post-transfection assessed by Western Blot. (D) Expression of ATP7B in each group at each time-point following transplantation in vivo. *p<0.05 compared with the saline group, &p<0.05 compared with IRP and RT group, respectively.

Mentions: At 48-hours post-transduction, nearly 100% of cells were transduced (figures 2A and 2B). To analyze the functional activity of BM-MSCs, 5 days after transduction, total proteins extracted from MSCsATP7B and MSCsGFP were analyzed by Western Blots. The protein levels of ATP7B were markedly enhanced in MSCsATP7B (figure 2C).


Transplantation of ATP7B-transduced bone marrow mesenchymal stem cells decreases copper overload in rats.

Chen S, Shao C, Dong T, Chai H, Xiong X, Sun D, Zhang L, Yu Y, Wang P, Cheng F - PLoS ONE (2014)

Transfection efficacy, expression of ATP7B post-transfection and post-transplantation.(A) Transfection efficacy of pRRL.PPT.SF.ATP7B.i2GFPBsd.pre in BM-MSCs. (B) Transfection efficacy of pRRL.PPT.SF.i2GFPpre in BM-MSCs. Transfected cells were observed under both a fluorescence microscope and a regular microscope (×100); (C) Expression of ATP7B in BM-MSCs post-transfection assessed by Western Blot. (D) Expression of ATP7B in each group at each time-point following transplantation in vivo. *p<0.05 compared with the saline group, &p<0.05 compared with IRP and RT group, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222898&req=5

pone-0111425-g002: Transfection efficacy, expression of ATP7B post-transfection and post-transplantation.(A) Transfection efficacy of pRRL.PPT.SF.ATP7B.i2GFPBsd.pre in BM-MSCs. (B) Transfection efficacy of pRRL.PPT.SF.i2GFPpre in BM-MSCs. Transfected cells were observed under both a fluorescence microscope and a regular microscope (×100); (C) Expression of ATP7B in BM-MSCs post-transfection assessed by Western Blot. (D) Expression of ATP7B in each group at each time-point following transplantation in vivo. *p<0.05 compared with the saline group, &p<0.05 compared with IRP and RT group, respectively.
Mentions: At 48-hours post-transduction, nearly 100% of cells were transduced (figures 2A and 2B). To analyze the functional activity of BM-MSCs, 5 days after transduction, total proteins extracted from MSCsATP7B and MSCsGFP were analyzed by Western Blots. The protein levels of ATP7B were markedly enhanced in MSCsATP7B (figure 2C).

Bottom Line: IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B.This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all).Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery Ward of General Surgery, The Affiliated Wuhu No. 2 People's Hospital of Wannan Medical College, Wuhu, Anhui Province, China.

ABSTRACT

Background: Recent studies have demonstrated that transplantation of ATP7B-transduced hepatocytes ameliorates disease progression in LEC (Long-Evans Cinnamon) rats, a model of Wilson's disease (WD). However, the inability of transplanted cells to proliferate in a normal liver hampers long-term treatment. In the current study, we investigated whether transplantation of ATP7B-transduced bone marrow mesenchymal stem cells (BM-MSCs) could decrease copper overload in LEC rats.

Materials and methods: The livers of LEC rats were preconditioned with radiation (RT) and/or ischemia-reperfusion (IRP) before portal vein infusion of ATP7B-transduced MSCs (MSCsATP7B). The volumes of MSCsATP7B or saline injected as controls were identical. The expression of ATP7B was analyzed by real-time quantitative polymerase chain reaction (RT-PCR) at 4, 12 and 24 weeks post-transplantation. MSCATP7B repopulation, liver copper concentrations, serum ceruloplasmin levels, and alanine transaminase (ALT) and aspartate transaminase (AST) levels were also analyzed at each time-point post-transplantation.

Results: IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B. This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all). Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).

Conclusions: These results demonstrate that transplantation of MSCsATP7B into IRP-plus-RT preconditioned LEC rats decreased copper overload and was associated with an increase in MSC engraftment and repopulation.

Show MeSH
Related in: MedlinePlus