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Gene Therapy Corrects Mitochondrial Dysfunction in Hematopoietic Progenitor Cells and Fibroblasts from Coq9R239X Mice.

Barriocanal-Casado E, Cueto-Ureña C, Benabdellah K, Gutiérrez-Guerrero A, Cobo M, Hidalgo-Gutiérrez A, Rodríguez-Sevilla JJ, Martín F, López LC - PLoS ONE (2016)

Bottom Line: A group of target diseases for these therapeutic strategies are mitochondrial encephalopathies due to mutations in nuclear DNA genes.Ectopic over-expression of Coq9 in both cell types restored the CoQ biosynthetic pathway and mitochondrial function, improving the fitness of the transduced cells.These results show the potential of the CCoq9WP lentiviral vector as a tool for gene therapy to treat mitochondrial encephalopathies.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.

ABSTRACT
Recent clinical trials have shown that in vivo and ex vivo gene therapy strategies can be an option for the treatment of several neurological disorders. Both strategies require efficient and safe vectors to 1) deliver the therapeutic gene directly into the CNS or 2) to genetically modify stem cells that will be used as Trojan horses for the systemic delivery of the therapeutic protein. A group of target diseases for these therapeutic strategies are mitochondrial encephalopathies due to mutations in nuclear DNA genes. In this study, we have developed a lentiviral vector (CCoq9WP) able to overexpress Coq9 mRNA and COQ9 protein in mouse embryonic fibroblasts (MEFs) and hematopoietic progenitor cells (HPCs) from Coq9R239X mice, an animal model of mitochondrial encephalopathy due to primary Coenzyme Q (CoQ) deficiency. Ectopic over-expression of Coq9 in both cell types restored the CoQ biosynthetic pathway and mitochondrial function, improving the fitness of the transduced cells. These results show the potential of the CCoq9WP lentiviral vector as a tool for gene therapy to treat mitochondrial encephalopathies.

No MeSH data available.


Related in: MedlinePlus

Transduction with CCoq9WP vector restores mitochondrial respiration and enhances fitness in MEFs from Coq9R239X mice.Oxygen consumption rate (OCR) profile (A), spare respiratory capacity (B) and cell index (C) in MEFs. Vertical arrows indicate the time of addition of oligomycin (olyg), FCCP and rotenone/antymicin (Rot/Ant). 5×104 cells were plated in each well. Data are expressed as mean ± SD. *P < 0.05, Coq9R239X and Coq9R239X-CCoq9WP cells versus Coq9+/+ cells; ++P < 0.01, Coq9R239X-CCoq9WP cells versus Coq9R239X cells; (one-way ANOVA with a Tukey's post hoc test; n = 4–6 for each group).
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pone.0158344.g005: Transduction with CCoq9WP vector restores mitochondrial respiration and enhances fitness in MEFs from Coq9R239X mice.Oxygen consumption rate (OCR) profile (A), spare respiratory capacity (B) and cell index (C) in MEFs. Vertical arrows indicate the time of addition of oligomycin (olyg), FCCP and rotenone/antymicin (Rot/Ant). 5×104 cells were plated in each well. Data are expressed as mean ± SD. *P < 0.05, Coq9R239X and Coq9R239X-CCoq9WP cells versus Coq9+/+ cells; ++P < 0.01, Coq9R239X-CCoq9WP cells versus Coq9R239X cells; (one-way ANOVA with a Tukey's post hoc test; n = 4–6 for each group).

Mentions: Skin fibroblasts of patients with primary CoQ10 deficiency show reduced activities of CoQ-dependent mitochondrial complexes and decreased levels of ATP [16, 19–23]. Similarly, isolated mitochondria from tissues of Coq9R239X show a reduction in mitochondrial respiration [3, 4]. Therefore, we next investigated whether the increase of CoQ9 induces functional changes in mitochondrial bioenergetics. We assessed the mitochondrial respiration in MEFs by SeaHorse Analyzer. Coq9R239X MEFs show a decrease in the global oxygen consumption rate and in the spare respiratory capacity (Fig 5A and 5B). Both alterations were normalized after transduction of Coq9R239X MEFs with the CCoq9WP LV (Fig 5A and 5B). As a consequence, cell growth was increased in CCoq9WP-transduced Coq9R239X compared to untransduced Coq9R239X MEFs and also to wild-type MEFs (Fig 5C). The improved cell growth was restricted to the first hours after plating, indicating that over-expression of COQ9 could improve adhesion of MEFs to plastic and this could favor initial growth (see green square in Fig 5C). After that moment, the growth rate was similar in the three experimental groups (Fig 5C).


Gene Therapy Corrects Mitochondrial Dysfunction in Hematopoietic Progenitor Cells and Fibroblasts from Coq9R239X Mice.

Barriocanal-Casado E, Cueto-Ureña C, Benabdellah K, Gutiérrez-Guerrero A, Cobo M, Hidalgo-Gutiérrez A, Rodríguez-Sevilla JJ, Martín F, López LC - PLoS ONE (2016)

Transduction with CCoq9WP vector restores mitochondrial respiration and enhances fitness in MEFs from Coq9R239X mice.Oxygen consumption rate (OCR) profile (A), spare respiratory capacity (B) and cell index (C) in MEFs. Vertical arrows indicate the time of addition of oligomycin (olyg), FCCP and rotenone/antymicin (Rot/Ant). 5×104 cells were plated in each well. Data are expressed as mean ± SD. *P < 0.05, Coq9R239X and Coq9R239X-CCoq9WP cells versus Coq9+/+ cells; ++P < 0.01, Coq9R239X-CCoq9WP cells versus Coq9R239X cells; (one-way ANOVA with a Tukey's post hoc test; n = 4–6 for each group).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920430&req=5

pone.0158344.g005: Transduction with CCoq9WP vector restores mitochondrial respiration and enhances fitness in MEFs from Coq9R239X mice.Oxygen consumption rate (OCR) profile (A), spare respiratory capacity (B) and cell index (C) in MEFs. Vertical arrows indicate the time of addition of oligomycin (olyg), FCCP and rotenone/antymicin (Rot/Ant). 5×104 cells were plated in each well. Data are expressed as mean ± SD. *P < 0.05, Coq9R239X and Coq9R239X-CCoq9WP cells versus Coq9+/+ cells; ++P < 0.01, Coq9R239X-CCoq9WP cells versus Coq9R239X cells; (one-way ANOVA with a Tukey's post hoc test; n = 4–6 for each group).
Mentions: Skin fibroblasts of patients with primary CoQ10 deficiency show reduced activities of CoQ-dependent mitochondrial complexes and decreased levels of ATP [16, 19–23]. Similarly, isolated mitochondria from tissues of Coq9R239X show a reduction in mitochondrial respiration [3, 4]. Therefore, we next investigated whether the increase of CoQ9 induces functional changes in mitochondrial bioenergetics. We assessed the mitochondrial respiration in MEFs by SeaHorse Analyzer. Coq9R239X MEFs show a decrease in the global oxygen consumption rate and in the spare respiratory capacity (Fig 5A and 5B). Both alterations were normalized after transduction of Coq9R239X MEFs with the CCoq9WP LV (Fig 5A and 5B). As a consequence, cell growth was increased in CCoq9WP-transduced Coq9R239X compared to untransduced Coq9R239X MEFs and also to wild-type MEFs (Fig 5C). The improved cell growth was restricted to the first hours after plating, indicating that over-expression of COQ9 could improve adhesion of MEFs to plastic and this could favor initial growth (see green square in Fig 5C). After that moment, the growth rate was similar in the three experimental groups (Fig 5C).

Bottom Line: A group of target diseases for these therapeutic strategies are mitochondrial encephalopathies due to mutations in nuclear DNA genes.Ectopic over-expression of Coq9 in both cell types restored the CoQ biosynthetic pathway and mitochondrial function, improving the fitness of the transduced cells.These results show the potential of the CCoq9WP lentiviral vector as a tool for gene therapy to treat mitochondrial encephalopathies.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Fisiología, Facultad de Medicina, Universidad de Granada, Granada, Spain.

ABSTRACT
Recent clinical trials have shown that in vivo and ex vivo gene therapy strategies can be an option for the treatment of several neurological disorders. Both strategies require efficient and safe vectors to 1) deliver the therapeutic gene directly into the CNS or 2) to genetically modify stem cells that will be used as Trojan horses for the systemic delivery of the therapeutic protein. A group of target diseases for these therapeutic strategies are mitochondrial encephalopathies due to mutations in nuclear DNA genes. In this study, we have developed a lentiviral vector (CCoq9WP) able to overexpress Coq9 mRNA and COQ9 protein in mouse embryonic fibroblasts (MEFs) and hematopoietic progenitor cells (HPCs) from Coq9R239X mice, an animal model of mitochondrial encephalopathy due to primary Coenzyme Q (CoQ) deficiency. Ectopic over-expression of Coq9 in both cell types restored the CoQ biosynthetic pathway and mitochondrial function, improving the fitness of the transduced cells. These results show the potential of the CCoq9WP lentiviral vector as a tool for gene therapy to treat mitochondrial encephalopathies.

No MeSH data available.


Related in: MedlinePlus