Isolation of transcriptomal changes attributable to LHON mutations and the cybridization process.
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.
Danielson
Steven R
SR
Department of Molecular Biosciences, University of California Davis, Davis, CA 95616, USA.
Carelli
Valerio
V
Tan
Guolin
G
Martinuzzi
Andrea
A
Schapira
Anthony H V
AH
Savontaus
Marja-Liisa
ML
Cortopassi
Gino A
GA
D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004272:DNA, Mitochondrial / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells; D020411:Oligonucleotide Array Sequence Analysis; D029242:Optic Atrophy, Hereditary, Leber / Q000235:genetics* / Q000378:metabolism; D017354:Point Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction; D013012:Sorbitol / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D004272:DNA, Mitochondrial; D013012:Sorbitol; D000449:Aldehyde Reductase