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Translation inhibition corrects aberrant localization of mutant alanine-glyoxylate aminotransferase: possible therapeutic approach for hyperoxaluria

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Abstract

Primary hyperoxaluria type 1 is a severe kidney stone disease caused by abnormalities of the peroxisomal alanine-glyoxylate aminotransferase (AGT). The most frequent mutation G170R results in aberrant mitochondrial localization of the active enzyme. To evaluate the population of peroxisome-localized AGT, we developed a quantitative Glow-AGT assay based on the self-assembly split-GFP approach and used it to identify drugs that can correct mislocalization of the mutant protein. In line with previous reports, the Glow-AGT assay showed that mitochondrial transport inhibitors DECA and monensin increased peroxisomal localization of the mutant. Here, we demonstrate that prolonged treatment with the translation elongation inhibitor emetine, a medicinal alkaloid used in treatment of amoebiasis, corrected G170R-AGT mislocalization. Furthermore, emetine reduced the augmented oxalate level in culture media of patient-derived hepatocytes bearing the G170R mutation. A distinct translation inhibitor GC7 had a similar effect on the mutant Glow-AGT relocalization indicating that mild translation inhibition is a promising therapeutic approach for primary hyperoxaluria type 1 caused by AGT misfolding/mistargeting.

Key messages

• There is no effective conservative treatment to decrease oxalate production in PH1 patients.

• Chemical chaperones rescue mislocalization of mutant AGT and reduce oxalate levels.

• We have developed an assay for precise monitoring of the peroxisomal AGT.

• Inhibition of translation by emetine reroutes the mutant protein to peroxisome.

• Mild translation inhibition is a promising cure for conformational disorders.

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Acknowledgements

We thank Stephanie Cabantous, Sonia Fargue, and Gill Rumsby for providing vectors and antibodies; Ellen Broide, Ruth Bargal, Rachel Cohen, Bruria Hirsh, and Anatoly Kustanovich for technical assistance; and Maya Schuldiner and Einat Zalckvar for fruitful discussions.

Funding

This work was funded by the GIF Research Grant No. I-1216-328.2/2012.

Author information

Author notes

  1. Roman Lyakhovetsky

    Present address: Medical Scientific Unit, Teva Pharmaceutical Industries, Petah Tikva, Israel

  2. Ruth Belostotsky and Roman Lyakhovetsky contributed equally to this work.

Authors and Affiliations

  1. Division of Pediatric Nephrology, Shaare Zedek Medical Center, Shmuel Bait Street, 91031, Jerusalem, Israel

    Ruth Belostotsky, Roman Lyakhovetsky, Fanny Shkedy, Shimrit Tzvi-Behr, Roi Bar & Yaacov Frishberg

  2. Department of Molecular Biology, Ariel University, Ariel, Israel

    Michael Y. Sherman

  3. Department of Pediatrics, University Medical Center, Bonn, Germany

    Bernd Hoppe

  4. Institute of Human Genetics, Cologne, Germany

    Björn Reusch & Bodo B. Beck

  5. Center for Molecular Medicine Cologne, Cologne, Germany

    Björn Reusch & Bodo B. Beck

Authors
  1. Ruth Belostotsky
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  2. Roman Lyakhovetsky
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Contributions

RB, YF, BB, and MS—designing research studies; RB, RL, BB, FS, STB, and RBar—conducting experiments; RB, RL, and BH—acquiring data; RB and RL—analyzing data; BB—providing reagents; and RB and YF—writing the manuscript.

Corresponding author

Correspondence to Ruth Belostotsky.

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The authors declare that they have no conflict of interest.

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Belostotsky, R., Lyakhovetsky, R., Sherman, M.Y. et al. Translation inhibition corrects aberrant localization of mutant alanine-glyoxylate aminotransferase: possible therapeutic approach for hyperoxaluria. J Mol Med 96, 621–630 (2018). https://doi.org/10.1007/s00109-018-1651-8

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  • DOI: https://doi.org/10.1007/s00109-018-1651-8

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