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Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells

Fabry disease (FD) is caused by mutations in the GLA gene that encodes lysosomal α-galactosidase-A (α-gal-A). A number of pathogenic mechanisms have been proposed and these include loss of mitochondrial respiratory chain activity. For FD, gene therapy is beginning to be applied as a treatment. In vi...

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Autores principales: Lambert, Jonathan R. A., Howe, Steven J., Rahim, Ahad A., Burke, Derek G., Heales, Simon J. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770804/
https://www.ncbi.nlm.nih.gov/pubmed/31491876
http://dx.doi.org/10.3390/ijms20184349
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author Lambert, Jonathan R. A.
Howe, Steven J.
Rahim, Ahad A.
Burke, Derek G.
Heales, Simon J. R.
author_facet Lambert, Jonathan R. A.
Howe, Steven J.
Rahim, Ahad A.
Burke, Derek G.
Heales, Simon J. R.
author_sort Lambert, Jonathan R. A.
collection PubMed
description Fabry disease (FD) is caused by mutations in the GLA gene that encodes lysosomal α-galactosidase-A (α-gal-A). A number of pathogenic mechanisms have been proposed and these include loss of mitochondrial respiratory chain activity. For FD, gene therapy is beginning to be applied as a treatment. In view of the loss of mitochondrial function reported in FD, we have considered here the impact of loss of mitochondrial respiratory chain activity on the ability of a GLA lentiviral vector to increase cellular α-gal-A activity and participate in cross correction. Jurkat cells were used in this study and were exposed to increasing viral copies. Intracellular and extracellular enzyme activities were then determined; this in the presence or absence of the mitochondrial complex I inhibitor, rotenone. The ability of cells to take up released enzyme was also evaluated. Increasing transgene copies was associated with increasing intracellular α-gal-A activity but this was associated with an increase in Km. Release of enzyme and cellular uptake was also demonstrated. However, in the presence of rotenone, enzyme release was inhibited by 37%. Excessive enzyme generation may result in a protein with inferior kinetic properties and a background of compromised mitochondrial function may impair the cross correction process.
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spelling pubmed-67708042019-10-30 Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells Lambert, Jonathan R. A. Howe, Steven J. Rahim, Ahad A. Burke, Derek G. Heales, Simon J. R. Int J Mol Sci Article Fabry disease (FD) is caused by mutations in the GLA gene that encodes lysosomal α-galactosidase-A (α-gal-A). A number of pathogenic mechanisms have been proposed and these include loss of mitochondrial respiratory chain activity. For FD, gene therapy is beginning to be applied as a treatment. In view of the loss of mitochondrial function reported in FD, we have considered here the impact of loss of mitochondrial respiratory chain activity on the ability of a GLA lentiviral vector to increase cellular α-gal-A activity and participate in cross correction. Jurkat cells were used in this study and were exposed to increasing viral copies. Intracellular and extracellular enzyme activities were then determined; this in the presence or absence of the mitochondrial complex I inhibitor, rotenone. The ability of cells to take up released enzyme was also evaluated. Increasing transgene copies was associated with increasing intracellular α-gal-A activity but this was associated with an increase in Km. Release of enzyme and cellular uptake was also demonstrated. However, in the presence of rotenone, enzyme release was inhibited by 37%. Excessive enzyme generation may result in a protein with inferior kinetic properties and a background of compromised mitochondrial function may impair the cross correction process. MDPI 2019-09-05 /pmc/articles/PMC6770804/ /pubmed/31491876 http://dx.doi.org/10.3390/ijms20184349 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lambert, Jonathan R. A.
Howe, Steven J.
Rahim, Ahad A.
Burke, Derek G.
Heales, Simon J. R.
Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title_full Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title_fullStr Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title_full_unstemmed Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title_short Inhibition of Mitochondrial Complex I Impairs Release of α-Galactosidase by Jurkat Cells
title_sort inhibition of mitochondrial complex i impairs release of α-galactosidase by jurkat cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770804/
https://www.ncbi.nlm.nih.gov/pubmed/31491876
http://dx.doi.org/10.3390/ijms20184349
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