Mitochondrial Mutations Research Project
Mitochondria: The “Powerhouse” of the Cell
Mitochondria provide energy for the cell by synthesizing energy in the form of high energy bonds. This energy synthesis occurs through a process called oxidative phosphorylation in which respiratory enzymes in mitochondria convert a molecule called adenosine diphosphate (ADP) into the energy currency of the cell, ATP. One interesting feature of mitochondria is that they contain their own DNA (mtDNA). As cells and mitochondria have co-evolved, most of this genetic information has been transferred to the nucleus, leaving only thirteen protein-encoding genes in the mtDNA.
The Problem: Energy Production Leads to mtDNA Mutations Accumulating Over Time
The Solution: Allotopic Expression of Mitochondrial Genes
SRF’s strategic approach to this problem is to engineer a way to let mitochondria keep producing energy normally, even after mitochondrial mutations have occurred. Although damage to mitochondrial DNA is inevitable so long as it is housed in the mitochondria, the harmful effects of mitochondrial mutations can be bypassed by engineering backup copies of the thirteen protein-encoding genes and housing the copies instead in the nucleus of the cell. These allotopic gene copies could continue to provide the necessary proteins even when mutations have compromised the mtDNA’s ability to do so. Moreover, the nuclear gene copies would be better shielded from damaging toxins and better maintained by DNA repair machinery. Since the majority of mitochondrial proteins are naturally nuclear-encoded, the natural mechanism to deliver the allotopically-expressed genes to the mitochondria can be co-opted.
Overcoming Obstacles to the Allotopic Expression Strategy
Previous attempts at allotopic expression have experienced only limited success due to problems with import of the proteins into the mitochondria, which were likely caused by the hydrophobic nature of the proteins. In addition to being ineffective, incomplete protein import is likely toxic to the mitochondrion and ultimately to the cell. To overcome these problems and achieve optimal mitochondrial import, SRF plans to use a co-translational import strategy. This strategy has been successfully employed by Professor Corral-Debrinski in work previously funded by SENS Research Foundation. The improvement that Dr. Corral-Debrinski has pioneered is tagging the RNA of the genes with sequences that not only target the proteins to the mitochondria but also direct the RNA to the mitochondrial surface before it is translated into protein. This approach prevents the convoluted folding of these proteins during translation in the watery environment of the cytosol, improving the efficiency of protein import.
Therapeutic Significance of the MitoSENS Project
Using mitochondrial gene therapy as a strategy to correct mitochondrial dysfunction has many advantages. In theory, such gene therapy could be used both to prevent and to correct the effects of mitochondrial mutations. Another exciting potential is that any therapy we develop could, in principle, be used to treat any of the known diseases of the mitochondria, such as LHON (Leber Hereditary Optic Neuropathy), Leigh syndrome, and NARP (Neurogenic muscle weakness, Ataxia, and Retinitis Pigmentosa), all of which are debilitating diseases. Indeed, allotopic expression is already being tested in human clinical trials to treat LHON. Treatment of these known and well-characterized diseases is a licensable therapeutic indication under current regulation, which therefore constitutes an entry point for the first human uses of mitochondrial gene therapies that emerge from our work.