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In vitro supplementation with dAMP/dGMP leads to partial restoration of mtDNA levels in mitochondrial depletion syndromes

In vitro supplementation with dAMP/dGMP leads to partial restoration of mtDNA levels in mitochondrial depletion syndromes
In vitro supplementation with dAMP/dGMP leads to partial restoration of mtDNA levels in mitochondrial depletion syndromes
Mitochondrial DNA depletion syndrome, a frequent cause of childhood (hepato)encephalomyopathies, is
defined as a reduction of mitochondrial DNA copy number related to nuclear DNA. It was previously
shown that mtDNA depletion can be prevented by dAMP/dGMP supplementation in deoxyguanosine
kinase-deficient fibroblasts. We investigated myotubes of patients diagnosed with mtDNA depletion carrying
pathogenic mutations in DGUOK, POLG1 (Alpers syndrome) and TYMP. Differentiating myotubes of all
patients and controls were supplemented with different doses of dAMP/dGMP or dAMP/dGMP/dCMP in
TYMP deficiency, and analysed for mtDNA/nDNA ratio and for cytochrome c oxidase (COX) activity. Serum
deprivation and myotube formation triggered a decrease in mtDNA copy number in DGUOK or POLG1
deficient myotubes, but not in TYMP deficiency and healthy controls. Supplementation with dAMP/dGMP
leads to a significant and reproducible rescue of mtDNA depletion in DGUOK deficiency. POLG1 deficient
myotubes also showed a mild, not significant increase in mtDNA copy number. MtDNA depletion did not
result in deficient COX staining in DGUOK and POLG1-deficient myotubes. Treatment with ethidium bromide
resulted in very severe depletion and absence of COX staining in all cell types, and no recovery was observed
after supplementation with dAMP/dGMP. We show that supplementation with dAMP/dGMP increases mtDNA
copy number significantly in DGUOK deficient myotubes and, leads to a mild, non-significant improvement of
mtDNA depletion in POLG1 deficiency. No adverse effect on mtDNA copy number was observed on high-dose
supplementation in vitro. Further studies are needed to determine possible therapeutic implications of dAMP/
dGMP supplementation for DGUOK deficiency in vivo.
1590-1599
Bulst, Stefanie
33f45952-4959-48bc-9986-227087693bd2
Abicht, Angela
8c9ac9fe-eb9f-40b2-8167-898c047532d1
Holinski-Feder, Elke
a11466cc-d92e-4398-89e2-d90d439c05c1
Mueller-Ziermann, Solvig
fbd3d8b6-f978-45df-9731-f881fdcb8383
Koehler, Udo
936dd5bc-4282-494b-b88c-2480ad280054
Thirion, Christian
bd06599d-28c0-4e03-a624-0cdd03544497
Walter, Maggie C.
005e23a3-d97d-47ac-bd02-9d3f0650fe05
Stewart, Joanna D.
e1ec9784-39cc-48ed-9f4f-2a05d25f2106
Chinnery, Patrick F.
a39c5ef7-0cac-401c-a3fb-2e0e2626e6dc
Lochmueller, Hanns
3681462f-af40-42e1-9714-ac723f7ee42d
Horvath, Rita
e41c9ffa-b0a0-4ebb-a44a-b98a2c6c23cd
Bulst, Stefanie
33f45952-4959-48bc-9986-227087693bd2
Abicht, Angela
8c9ac9fe-eb9f-40b2-8167-898c047532d1
Holinski-Feder, Elke
a11466cc-d92e-4398-89e2-d90d439c05c1
Mueller-Ziermann, Solvig
fbd3d8b6-f978-45df-9731-f881fdcb8383
Koehler, Udo
936dd5bc-4282-494b-b88c-2480ad280054
Thirion, Christian
bd06599d-28c0-4e03-a624-0cdd03544497
Walter, Maggie C.
005e23a3-d97d-47ac-bd02-9d3f0650fe05
Stewart, Joanna D.
e1ec9784-39cc-48ed-9f4f-2a05d25f2106
Chinnery, Patrick F.
a39c5ef7-0cac-401c-a3fb-2e0e2626e6dc
Lochmueller, Hanns
3681462f-af40-42e1-9714-ac723f7ee42d
Horvath, Rita
e41c9ffa-b0a0-4ebb-a44a-b98a2c6c23cd

Bulst, Stefanie, Abicht, Angela, Holinski-Feder, Elke, Mueller-Ziermann, Solvig, Koehler, Udo, Thirion, Christian, Walter, Maggie C., Stewart, Joanna D., Chinnery, Patrick F., Lochmueller, Hanns and Horvath, Rita (2009) In vitro supplementation with dAMP/dGMP leads to partial restoration of mtDNA levels in mitochondrial depletion syndromes. Human Molecular Genetics, 18 (9), 1590-1599. (doi:10.1093/hmg/ddp074). (PMID:19221117)

Record type: Article

Abstract

Mitochondrial DNA depletion syndrome, a frequent cause of childhood (hepato)encephalomyopathies, is
defined as a reduction of mitochondrial DNA copy number related to nuclear DNA. It was previously
shown that mtDNA depletion can be prevented by dAMP/dGMP supplementation in deoxyguanosine
kinase-deficient fibroblasts. We investigated myotubes of patients diagnosed with mtDNA depletion carrying
pathogenic mutations in DGUOK, POLG1 (Alpers syndrome) and TYMP. Differentiating myotubes of all
patients and controls were supplemented with different doses of dAMP/dGMP or dAMP/dGMP/dCMP in
TYMP deficiency, and analysed for mtDNA/nDNA ratio and for cytochrome c oxidase (COX) activity. Serum
deprivation and myotube formation triggered a decrease in mtDNA copy number in DGUOK or POLG1
deficient myotubes, but not in TYMP deficiency and healthy controls. Supplementation with dAMP/dGMP
leads to a significant and reproducible rescue of mtDNA depletion in DGUOK deficiency. POLG1 deficient
myotubes also showed a mild, not significant increase in mtDNA copy number. MtDNA depletion did not
result in deficient COX staining in DGUOK and POLG1-deficient myotubes. Treatment with ethidium bromide
resulted in very severe depletion and absence of COX staining in all cell types, and no recovery was observed
after supplementation with dAMP/dGMP. We show that supplementation with dAMP/dGMP increases mtDNA
copy number significantly in DGUOK deficient myotubes and, leads to a mild, non-significant improvement of
mtDNA depletion in POLG1 deficiency. No adverse effect on mtDNA copy number was observed on high-dose
supplementation in vitro. Further studies are needed to determine possible therapeutic implications of dAMP/
dGMP supplementation for DGUOK deficiency in vivo.

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Published date: 16 February 2009
Organisations: Centre for Biological Sciences

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Local EPrints ID: 355880
URI: https://eprints.soton.ac.uk/id/eprint/355880
PURE UUID: 205811c7-c9e3-4d78-90b0-55364e1627e9

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Date deposited: 04 Sep 2013 17:19
Last modified: 18 Jul 2017 03:43

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Contributors

Author: Stefanie Bulst
Author: Angela Abicht
Author: Elke Holinski-Feder
Author: Solvig Mueller-Ziermann
Author: Udo Koehler
Author: Christian Thirion
Author: Maggie C. Walter
Author: Joanna D. Stewart
Author: Patrick F. Chinnery
Author: Hanns Lochmueller
Author: Rita Horvath

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