Abstract
Nonsense mutations, resulting in a premature stop codon in the open reading frame of mRNAs are responsible for thousands of inherited diseases. Readthrough of premature stop codons by small molecule drugs has emerged as a promising therapeutic approach to treat disorders resulting from premature termination of translation. The aminoglycoside antibiotics are a class of molecule known to promote readthrough at premature termination codons. Gentamicin consists of a mixture of major and minor aminoglycoside components. Here, we investigated the readthrough activities of the individual components and show that each of the four major gentamicin complex components representing 92–99% of the complex each had similar potency and activity to that of the complex itself. In contrast, a minor component (gentamicin X2) was found to be the most potent and active readthrough component in the gentamicin complex. The known oto- and nephrotoxicity associated with aminoglycosides preclude long-term use as readthrough agents. Thus, we evaluated the components of the gentamicin complex as well as the so-called “designer” aminoglycoside, NB124, for in vitro and in vivo safety. In cells, we observed that gentamicin X2 had a safety/readthrough ratio (cytotoxicity/readthrough potency) superior to that of gentamicin, G418 or NB124. In rodents, we observed that gentamicin X2 showed a safety profile that was superior to G418 overall including reduced nephrotoxicity. These results support further investigation of gentamicin X2 as a therapeutic readthrough agent.
Highlights
The presence of a nonsense mutation in the open reading frame of a gene leads to the introduction of a premature termination codon in the mRNA that results in the production of a truncated-nonfunctional protein product
Gentamicin X2, a minor component of the gentamicin complex, is a potent readthrough compound. Because it is approved for use as an antibiotic, gentamicin has been investigated as a potential readthrough therapeutic in many different nonsense mutation disease models [29]
In a search to identify therapeutic aminoglycosides with reduced toxicity and increased readthrough potency and activity, we evaluated all of the commercially available gentamicin components and compared their biological activity to that of G418 and the so-called “designer” aminoglycosides NB84 and NB124
Summary
The presence of a nonsense mutation in the open reading frame of a gene leads to the introduction of a premature termination codon in the mRNA that results in the production of a truncated-nonfunctional protein product. Nonsense mutations are the cause of approximately 11% of all genetic diseases [1]. Readthrough of premature termination codons to allow.
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