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A catalytically silent FAAH-1 variant drives anandamide transport in neurons

Nature Neuroscience volume 15, pages 64–69 (2012)Cite this article

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A Corrigendum to this article was published on 22 November 2013

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Abstract

The endocannabinoid anandamide is removed from the synaptic space by a selective transport system, expressed in neurons and astrocytes, that remains molecularly uncharacterized. Here we describe a partly cytosolic variant of the intracellular anandamide-degrading enzyme fatty acid amide hydrolase-1 (FAAH-1), termed FAAH-like anandamide transporter (FLAT), that lacked amidase activity but bound anandamide with low micromolar affinity and facilitated its translocation into cells. Known anandamide transport inhibitors, such as AM404 and OMDM-1, blocked these effects. We also identified a competitive antagonist of the interaction of anandamide with FLAT, the phthalazine derivative ARN272, that prevented anandamide internalization in vitro, interrupted anandamide deactivation in vivo and exerted profound analgesic effects in rodent models of nociceptive and inflammatory pain, which were mediated by CB1 cannabinoid receptors. The results identify FLAT as a critical molecular component of anandamide transport in neural cells and a potential target for therapeutic drugs.

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Figure 1: Structural properties of FLAT.
Figure 2: FLAT binds to anandamide and facilitates its transport into cells.
Figure 3: ARN272 is a competitive FLAT inhibitor.
Figure 4: ARN272 produces CB1-dependent antinociception in mice.

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  • 15 December 2011

    In the version of this article initially published, ref. 36 (given as Kaczowka, S.J. et al. Int. J. Biol. Macromol. 42, 62–67, 2008) was incorrect. The correct reference is Kaczocha, M., Glaser, S.T. & Deutsch, D.G. Identification of intracellular carriers for the endocannabinoid anandamide. Proc. Natl. Acad. Sci. USA 106, 6375–6380 (2009). The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank G. Colombano, J. Kim, S. Mandrup-Bertozzi, D. Thongkham, G. Moreno-Sanz and R. Scarpelli for help with experiments and discussions about the project. This research was supported by grants from the US National Institute on Drug Abuse and the US National Institute on Alcohol Abuse and Alcoholism (DA-012413 to D.P.) and the US National Institute on General Medicine (1R01GM093937-01 to W.R.). The support provided by the Agilent/University of California Irvine Analytical Discovery Facility is gratefully acknowledged.

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  1. Jin Fu and Giovanni Bottegoni: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology, University of California, Irvine, Irvine, California, USA

    Jin Fu, Ana Guijarro & Daniele Piomelli

  2. Drug Discovery and Development, Italian Institute of Technology, Genoa, Italy

    Giovanni Bottegoni, Oscar Sasso, Rosalia Bertorelli, Walter Rocchia, Matteo Masetti, Andrea Armirotti, Gianpiero Garau, Tiziano Bandiera, Angelo Reggiani, Andrea Cavalli & Daniele Piomelli

  3. Pharmaceutical Department, University of Parma, Parma, Italy

    Alessio Lodola & Marco Mor

  4. Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

    Andrea Cavalli

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Contributions

J.F. conducted the molecular biological, biochemical and pharmacological experiments in vitro and analyzed data. G.B. conducted the virtual ligand screening campaign. G.B., W.R., M. Masetti, A.L. and A.C. performed the computational studies. T.B. provided chemical expertise. O.S., R.B. and A.R. conducted the pharmacological experiments in vivo and analyzed data. A.G. measured lipid levels in vivo and analyzed data. A.A. and G.G. conducted analytical studies on purified recombinant FLAT. D.P. conceived and designed the experiments, oversaw the project and wrote the manuscript with assistance from J.F., G.B., W.R., M. Mor, A.L. and A.C.

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Correspondence to Daniele Piomelli.

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A patent was filed by the University of California, Irvine, and the Italian Institute of Technology on behalf of D.P., J.F., G.B. and A.C.

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Fu, J., Bottegoni, G., Sasso, O. et al. A catalytically silent FAAH-1 variant drives anandamide transport in neurons. Nat Neurosci 15, 64–69 (2012). https://doi.org/10.1038/nn.2986

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