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Self-sufficient control of urate homeostasis in mice by a synthetic circuit

Nature Biotechnology volume 28, pages 355–360 (2010)Cite this article

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Abstract

Synthetic biology has shown that the metabolic behavior of mammalian cells can be altered by genetic devices such as epigenetic and hysteretic switches1,2, timers and oscillators3,4, biocomputers5, hormone systems6 and heterologous metabolic shunts7. To explore the potential of such devices for therapeutic strategies, we designed a synthetic mammalian circuit to maintain uric acid homeostasis in the bloodstream, disturbance of which is associated with tumor lysis syndrome and gout8,9. This synthetic device consists of a modified Deinococcus radiodurans-derived protein that senses uric acids levels and triggers dose-dependent derepression of a secretion-engineered Aspergillus flavus urate oxidase that eliminates uric acid. In urate oxidase–deficient mice, which develop acute hyperuricemia, the synthetic circuit decreased blood urate concentration to stable sub-pathologic levels in a dose-dependent manner and reduced uric acid crystal deposits in the kidney. Synthetic gene-network devices providing self-sufficient control of pathologic metabolites represent molecular prostheses, which may foster advances in future gene- and cell-based therapies.

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Figure 1: Synthetic uric acid–responsive mammalian sensor circuit.
Figure 2: Validation of UREX-controlled SEAP expression in transgenic HEK-293 and urate oxidase–deficient mice.
Figure 3: Functional characterization of an engineered mammalian A. flavus–derived urate oxidase.
Figure 4: UREX-controlled smUox-mediated reduction of pathologic urate levels in mice.

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Acknowledgements

We thank W. Weber (Albert-Ludwigs-Universität Freiburg, Germany) for conceptual input and for providing pBluescript-mHucR, M. Gilet for skillful assistance with the animal study and Marcia Schoenberg for critical comments on the manuscript. This work was supported by the Swiss National Science Foundation (grant no. 31003A-126022) and in part by the EC Framework 7 (Persist).

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Authors and Affiliations

  1. Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland

    Christian Kemmer, Marc Gitzinger, Jörg Stelling & Martin Fussenegger

  2. Département Génie Biologique, Institut Universitaire de Technologie, IUTA, Villeurbanne Cedex, France

    Marie Daoud-El Baba

  3. Département de Médecine, Université de Fribourg, Fribourg, Switzerland

    Valentin Djonov

  4. Swiss Institute of Bioinformatics, ETH Zurich, Zurich, Switzerland

    Jörg Stelling

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  1. Christian Kemmer
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  4. Valentin Djonov
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  5. Jörg Stelling
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  6. Martin Fussenegger
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Contributions

C.K., J.S. and M.F. designed the project, analyzed results and wrote the manuscript. C.K., M.G., M.D.-E.B. and V.D. performed the experimental work. J.S. designed the mathematical model and performed simulations.

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Correspondence to Jörg Stelling or Martin Fussenegger.

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The authors declare no competing financial interests.

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Supplementary Figs. 1–13 and Supplementary Tables 1–6 and Supplementary Results (PDF 407 kb)

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Kemmer, C., Gitzinger, M., Daoud-El Baba, M. et al. Self-sufficient control of urate homeostasis in mice by a synthetic circuit. Nat Biotechnol 28, 355–360 (2010). https://doi.org/10.1038/nbt.1617

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