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Self-adjusting synthetic gene circuit for correcting insulin resistance

Nature Biomedical Engineering volume 1, Article number: 0005 (2017) Cite this article

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Abstract

Sophisticated genetic devices can be assembled to reprogram mammalian cell activities using tools from synthetic biology. Here, we demonstrate that a self-adjusting synthetic gene circuit can be designed to sense and reverse the insulin-resistance syndrome in different mouse models. By functionally rewiring the mitogen-activated protein kinase (MAPK) signalling pathway to produce MAPK-mediated activation of a hybrid transcription factor consisting of the tetracycline repressor, TetR, fused to the human ELK1-derived transactivation domain (TetR-Elk1), we assembled a synthetic insulin-sensitive transcription-control device that self-sufficiently distinguished between physiological and increased blood insulin levels and correspondingly fine-tuned the reversible expression of therapeutic transgenes from synthetic TetR-ELK1-specific promoters. In acute experimental hyperinsulinaemia, the synthetic insulin-sensing designer circuit reversed the insulin-resistance syndrome by coordinating expression of the insulin-sensitizing compound adiponectin. Engineering synthetic gene circuits to sense pathologic markers and coordinate the expression of therapeutic transgenes may provide opportunities for future gene- and cell-based treatments of multifactorial metabolic disorders.

In recent decades, obesity has resulted in an epidemic prevalence of type 2 diabetes, affecting more than 400 million individuals worldwide1. Obesity-induced insulin resistance results from complex metabolic and inflammatory changes and is the key etiologic defect of the metabolic syndrome, a cluster of clinical findings including hypertension and dyslipidaemia that increase the imminence of cardiovascular disorders and type 2 diabetes2,3. The physiological response to insulin resistance and impaired glucose metabolism is the continuous stimulation of insulin secretion, which initiates the gradual decline of pancreatic beta cells, leading to progressive development of type 2 diabetes and its acute and chronic complications4. Current insulin-resistance therapies, which focus on the reduction of body weight or the administration of insulin-sensitizing drugs such as glitazones, are ineffective or associated with important side effects2,5–7. Insulin resistance is inversely correlated with blood levels of adiponectin8–10, the most abundant adipose tissue-derived hormone with insulin-sensitizing properties11. Therefore, the development of drugs that increase blood adiponectin levels is currently being pursued for the treatment of insulin resistance and the associated cardiovascular risk factors10.

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Figure 1: Synthetic insulin-sensitizing designer circuit for the treatment of insulin resistance.
Figure 2: Synthetic insulin-inducible mammalian sensor circuit.
Figure 3: Self-sufficient insulin-sensor-based control of adiponectin expression in insulin-resistant ob/ob mice.
Figure 4: Self-sufficient insulin-sensor-based control of adiponectin expression in insulin-resistant DIO mice.
Figure 5: Long-term therapeutic efficacy of insulin-triggered adiponectin expression in insulin-resistant ob/ob mice.
Figure 6: Long-term therapeutic efficacy of insulin-triggered adiponectin expression in insulin-resistant DIO mice.

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Acknowledgements

We thank T. Abel for providing the pTetR-ELK1 (MKp37) plasmid, B. Geering for providing human serum from healthy individuals, Y. Lai for providing the DyLight 800-labelled goat anti-mouse IgG, B. M. Lang and L. Scheller for assistance with the statistical analyses and M. Daoud-El Baba for skilful assistance with the animal study. This work was supported by a European Research Council (ERC) advanced grant (no. 321381), the Cantons of Basel and the Swiss Confederation within the INTERREG IV A.20 tri-national research program and the Gutenberg Chair (awarded to M.F.). This work was also supported by the National Key Research and Development Program of China, Stem Cell and Translational Research (no. 2016YFA0100300), the National Natural Science Foundation of China (NSFC; nos 31470834, 31522017 and 31670869), the Science and Technology Commission of Shanghai Municipality (nos 15QA1401500 and 14JC1401700) and Thousand Youth Talents Plan (awarded to H.Y.).

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Author notes

  1. Haifeng Ye and Mingqi Xie: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, Basel, CH–4058, Switzerland

    Haifeng Ye, Mingqi Xie, Henryk Zulewski & Martin Fussenegger

  2. Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Dongchuan Road 500, Shanghai, 200241, China

    Haifeng Ye, Shuai Xue & Jianli Yin

  3. Département Génie Biologique, Institut Universitaire de Technologie, IUT, Villeurbanne Cedex, 69622, France

    Ghislaine Charpin-El Hamri

  4. Division of Endocrinology, Diabetes and Metabolism, University Hospital Basel, Petersgraben 4, Basel, CH–4031, Switzerland

    Henryk Zulewski

  5. Division of Endocrinology and Diabetes, Stadtspital Triemli, Birmensdorferstrasse 497, Zurich, CH–8063, Switzerland

    Henryk Zulewski

  6. University of Basel, Faculty of Science, Mattenstrasse 26, Basel, CH–4058, Switzerland

    Martin Fussenegger

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Contributions

H.Y., M.X., H.Z. and M.F. designed the project, analysed the results and wrote the manuscript. H.Y., M.X., G.H.E., S.X. and J.Y. performed the experimental work.

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Correspondence to Haifeng Ye or Martin Fussenegger.

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Ye, H., Xie, M., Xue, S. et al. Self-adjusting synthetic gene circuit for correcting insulin resistance. Nat Biomed Eng 1, 0005 (2017). https://doi.org/10.1038/s41551-016-0005

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