Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process

Steven R. Danielson, Valerio Carelli, Guolin Tan, Andrea Martinuzzi, Anthony H V Schapira, Marja Liisa Savontaus, Gino A Cortopassi

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Leber's hereditary optic neuropathy (LHON) is thought to be the most common disease resulting from mitochondrial DNA (mtDNA) point mutations, and transmitochondrial cytoplasmic hybrid (cybrid) cell lines are the most frequently used model for understanding the pathogenesis of mitochondrial disorders. We have used oligonucleotide microarrays and a novel study design based on shared transcripts to allocate transcriptomal changes into rho-zero-dependent, cybridization-dependent and LHON-dependent categories in these cells. The analysis indicates that the rho-zero process has the largest transcriptomal impact, followed by the cybridization process, and finally the LHON mutations. The transcriptomal impacts of the rho-zero and cybridization processes preferentially and significantly affect the mitochondrial compartment, causing upregulation of many transcripts involved in oxidative phosphorylation, presumably in response to the mtDNA depletion that occurs at the rho-zero step. Nine LHON-specific transcriptional alterations were shared among osteosarcoma cybrids and lymphoblasts bearing LHON mutations. Notably, the aldose reductase transcript was overexpressed in LHON cybrids and lymphoblasts. Aldose reductase is also overexpressed in diabetic retinopathy, leading to optic nerve and retinal complications. The LHON-specific increase in transcript level was confirmed by quantitative reverse transcription-polymerase chain reaction (RT-PCR), and a western blot confirmed a higher level of aldose reductase in mutant mitochondria. One product of aldose reductase is sorbitol, which has been linked to osmotic stress, oxidative stress and optic neuropathy, and sorbitol levels were increased in LHON cybrids. If these results are confirmed in patient tissues, aldose reductase inhibitors could have some therapeutic value for LHON.

Original languageEnglish (US)
Pages (from-to)1026-1037
Number of pages12
JournalBrain
Volume128
Issue number5
DOIs
StatePublished - May 2005

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Leber's Hereditary Optic Atrophy
Aldehyde Reductase
Mutation
Sorbitol
Mitochondrial DNA
Mitochondrial Diseases
Optic Nerve Diseases
Hybrid Cells
Oxidative Phosphorylation
Osmotic Pressure
Diabetic Retinopathy
Osteosarcoma
Optic Nerve
Oligonucleotide Array Sequence Analysis
Point Mutation
Reverse Transcription
Mitochondria
Oxidative Stress
Up-Regulation
Western Blotting

Keywords

  • Aldose reductase
  • Cybrid
  • Leber's hereditary optic neuropathy
  • Microarray
  • Rho-zero

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Danielson, S. R., Carelli, V., Tan, G., Martinuzzi, A., Schapira, A. H. V., Savontaus, M. L., & Cortopassi, G. A. (2005). Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process. Brain, 128(5), 1026-1037. https://doi.org/10.1093/brain/awh447

Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process. / Danielson, Steven R.; Carelli, Valerio; Tan, Guolin; Martinuzzi, Andrea; Schapira, Anthony H V; Savontaus, Marja Liisa; Cortopassi, Gino A.

In: Brain, Vol. 128, No. 5, 05.2005, p. 1026-1037.

Research output: Contribution to journalArticle

Danielson, SR, Carelli, V, Tan, G, Martinuzzi, A, Schapira, AHV, Savontaus, ML & Cortopassi, GA 2005, 'Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process', Brain, vol. 128, no. 5, pp. 1026-1037. https://doi.org/10.1093/brain/awh447
Danielson SR, Carelli V, Tan G, Martinuzzi A, Schapira AHV, Savontaus ML et al. Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process. Brain. 2005 May;128(5):1026-1037. https://doi.org/10.1093/brain/awh447
Danielson, Steven R. ; Carelli, Valerio ; Tan, Guolin ; Martinuzzi, Andrea ; Schapira, Anthony H V ; Savontaus, Marja Liisa ; Cortopassi, Gino A. / Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process. In: Brain. 2005 ; Vol. 128, No. 5. pp. 1026-1037.
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N2 - Leber's hereditary optic neuropathy (LHON) is thought to be the most common disease resulting from mitochondrial DNA (mtDNA) point mutations, and transmitochondrial cytoplasmic hybrid (cybrid) cell lines are the most frequently used model for understanding the pathogenesis of mitochondrial disorders. We have used oligonucleotide microarrays and a novel study design based on shared transcripts to allocate transcriptomal changes into rho-zero-dependent, cybridization-dependent and LHON-dependent categories in these cells. The analysis indicates that the rho-zero process has the largest transcriptomal impact, followed by the cybridization process, and finally the LHON mutations. The transcriptomal impacts of the rho-zero and cybridization processes preferentially and significantly affect the mitochondrial compartment, causing upregulation of many transcripts involved in oxidative phosphorylation, presumably in response to the mtDNA depletion that occurs at the rho-zero step. Nine LHON-specific transcriptional alterations were shared among osteosarcoma cybrids and lymphoblasts bearing LHON mutations. Notably, the aldose reductase transcript was overexpressed in LHON cybrids and lymphoblasts. Aldose reductase is also overexpressed in diabetic retinopathy, leading to optic nerve and retinal complications. The LHON-specific increase in transcript level was confirmed by quantitative reverse transcription-polymerase chain reaction (RT-PCR), and a western blot confirmed a higher level of aldose reductase in mutant mitochondria. One product of aldose reductase is sorbitol, which has been linked to osmotic stress, oxidative stress and optic neuropathy, and sorbitol levels were increased in LHON cybrids. If these results are confirmed in patient tissues, aldose reductase inhibitors could have some therapeutic value for LHON.

AB - Leber's hereditary optic neuropathy (LHON) is thought to be the most common disease resulting from mitochondrial DNA (mtDNA) point mutations, and transmitochondrial cytoplasmic hybrid (cybrid) cell lines are the most frequently used model for understanding the pathogenesis of mitochondrial disorders. We have used oligonucleotide microarrays and a novel study design based on shared transcripts to allocate transcriptomal changes into rho-zero-dependent, cybridization-dependent and LHON-dependent categories in these cells. The analysis indicates that the rho-zero process has the largest transcriptomal impact, followed by the cybridization process, and finally the LHON mutations. The transcriptomal impacts of the rho-zero and cybridization processes preferentially and significantly affect the mitochondrial compartment, causing upregulation of many transcripts involved in oxidative phosphorylation, presumably in response to the mtDNA depletion that occurs at the rho-zero step. Nine LHON-specific transcriptional alterations were shared among osteosarcoma cybrids and lymphoblasts bearing LHON mutations. Notably, the aldose reductase transcript was overexpressed in LHON cybrids and lymphoblasts. Aldose reductase is also overexpressed in diabetic retinopathy, leading to optic nerve and retinal complications. The LHON-specific increase in transcript level was confirmed by quantitative reverse transcription-polymerase chain reaction (RT-PCR), and a western blot confirmed a higher level of aldose reductase in mutant mitochondria. One product of aldose reductase is sorbitol, which has been linked to osmotic stress, oxidative stress and optic neuropathy, and sorbitol levels were increased in LHON cybrids. If these results are confirmed in patient tissues, aldose reductase inhibitors could have some therapeutic value for LHON.

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