Huntingtin (HTT)-lowering therapies hold promise to slow down neurodegeneration in Huntington's disease (HD). Here, we assessed the translatability and long-term durability of recombinant adeno-associated viral vector serotype 5 expressing a microRNA targeting human HTT (rAAV5-miHTT) administered by magnetic resonance imaging-guided convention-enhanced delivery in transgenic HD minipigs. rAAV5-miHTT (1.2 × 1013 vector genome (VG) copies per brain) was successfully administered into the striatum (bilaterally in caudate and putamen), using age-matched untreated animals as controls. Widespread brain biodistribution of vector DNA was observed, with the highest concentration in target (striatal) regions, thalamus, and cortical regions. Vector DNA presence and transgene expression were similar at 6 and 12 months after administration. Expression of miHTT strongly correlated with vector DNA, with a corresponding reduction of mutant HTT (mHTT) protein of more than 75% in injected areas, and 30 to 50% lowering in distal regions. Translational pharmacokinetic and pharmacodynamic measures in cerebrospinal fluid (CSF) were largely in line with the effects observed in the brain. CSF miHTT expression was detected up to 12 months, with CSF mHTT protein lowering of 25 to 30% at 6 and 12 months after dosing. This study demonstrates widespread biodistribution, strong and durable efficiency of rAAV5-miHTT in disease-relevant regions in a large brain, and the potential of using CSF analysis to determine vector expression and efficacy in the clinic.
- MeSH
- genetická terapie MeSH
- genetické vektory genetika MeSH
- Huntingtonova nemoc * genetika terapie MeSH
- lidé MeSH
- mikro RNA * metabolismus MeSH
- miniaturní prasata metabolismus MeSH
- modely nemocí na zvířatech MeSH
- prasata MeSH
- protein huntingtin genetika metabolismus MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Reactive metabolites of benzene 1,4-benzoquinone and 1,4-hydroquinone exert their toxic effects through covalent and/or oxidative damage to DNA and proteins. Since minipigs have been proposed as a suitable model species in toxicological and pharmacological research, the aim of this study was to explore mechanisms by which catechol, 1,4-hydroquinone and 1,4-benzoquinone destroy cytochrome P450 (P450) and induce oxidative stress in minipig liver microsomes. Our second goal was to assess the usefulness of minipig liver microsomes as a model system for the testing of the production of oxidative stress by clinically relevant quinone-containing compounds, e.g. anthracyclines. Of the three benzene metabolites tested, the highest P450 destruction was caused by 1,4-benzoquinone. This destructive effect did not correlate with the production of hydroxyl radicals as measured by ESR spin trapping which was the highest in samples containing 1,4-hydroquinone. Our results confirm previous findings that 1,4-benzoquinone exerts its effect mainly by direct attack on macromolecules while 1,4-hydroquinone rather stimulates the production of reactive oxygen species. Doxorubicin stimulated the production of hydroxyl radicals and the destruction of P450 similarly as 1,4-hydroquinone. Minipig liver microsomes should be further tested as a possibly suitable model system for the testing of potential modulators of the toxicity of doxorubicin.
- MeSH
- algoritmy MeSH
- antimetabolity antitumorózní toxicita MeSH
- benzen MeSH
- benzochinony metabolismus toxicita MeSH
- cytochrom P-450 CYP2E1 metabolismus MeSH
- cytochrom P-450 CYP3A metabolismus MeSH
- doxorubicin toxicita MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- elektronová paramagnetická rezonance MeSH
- financování organizované MeSH
- hydrochinony metabolismus toxicita MeSH
- hydroxylový radikál metabolismus MeSH
- inhibitory cytochromu P450 MeSH
- jaterní mikrozomy metabolismus účinky léků MeSH
- miniaturní prasata metabolismus MeSH
- NADP toxicita MeSH
- orchiektomie MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- prasata MeSH
- systém (enzymů) cytochromů P-450 MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- MeSH
- izoenzymy metabolismus MeSH
- jaterní mikrozomy enzymologie MeSH
- lidé MeSH
- miniaturní prasata metabolismus MeSH
- systém (enzymů) cytochromů P-450 metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
