CERT1 mutations perturb human development by disrupting sphingolipid homeostasis
Charlotte Gehin, et al.
Charlotte Gehin, et al.
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Research Article Cell biology Genetics

CERT1 mutations perturb human development by disrupting sphingolipid homeostasis

Abstract

Neural differentiation, synaptic transmission, and action potential propagation depend on membrane sphingolipids, whose metabolism is tightly regulated. Mutations in the ceramide transporter CERT (CERT1), which is involved in sphingolipid biosynthesis, are associated with intellectual disability, but the pathogenic mechanism remains obscure. Here, we characterize 31 individuals with de novo missense variants in CERT1. Several variants fall into a previously uncharacterized dimeric helical domain that enables CERT homeostatic inactivation, without which sphingolipid production goes unchecked. The clinical severity reflects the degree to which CERT autoregulation is disrupted, and inhibiting CERT pharmacologically corrects morphological and motor abnormalities in a Drosophila model of the disease, which we call ceramide transporter (CerTra) syndrome. These findings uncover a central role for CERT autoregulation in the control of sphingolipid biosynthetic flux, provide unexpected insight into the structural organization of CERT, and suggest a possible therapeutic approach for patients with CerTra syndrome.

Authors

Charlotte Gehin, Museer A. Lone, Winston Lee, Laura Capolupo, Sylvia Ho, Adekemi M. Adeyemi, Erica H. Gerkes, Alexander P.A. Stegmann, Estrella López-Martín, Eva Bermejo-Sánchez, Beatriz Martínez-Delgado, Christiane Zweier, Cornelia Kraus, Bernt Popp, Vincent Strehlow, Daniel Gräfe, Ina Knerr, Eppie R. Jones, Stefano Zamuner, Luciano A. Abriata, Vidya Kunnathully, Brandon E. Moeller, Anthony Vocat, Samuel Rommelaere, Jean-Philippe Bocquete, Evelyne Ruchti, Greta Limoni, Marine Van Campenhoudt, Samuel Bourgeat, Petra Henklein, Christian Gilissen, Bregje W. van Bon, Rolph Pfundt, Marjolein H. Willemsen, Jolanda H. Schieving, Emanuela Leonardi, Fiorenza Soli, Alessandra Murgia, Hui Guo, Qiumeng Zhang, Kun Xia, Christina R. Fagerberg, Christoph P. Beier, Martin J. Larsen, Irene Valenzuela, Paula Fernández-Álvarez, Shiyi Xiong, Robert Śmigiel, Vanesa López-González, Lluís Armengol, Manuela Morleo, Angelo Selicorni, Annalaura Torella, Moira Blyth, Nicola S. Cooper, Valerie Wilson, Renske Oegema, Yvan Herenger, Aurore Garde, Ange-Line Bruel, Frederic Tran Mau-Them, Alexis B.R. Maddocks, Jennifer M. Bain, Musadiq A. Bhat, Gregory Costain, Peter Kannu, Ashish Marwaha, Neena L. Champaigne, Michael J. Friez, Ellen B. Richardson, Vykuntaraju K. Gowda, Varunvenkat M. Srinivasan, Yask Gupta, Tze Y. Lim, Simone Sanna-Cherchi, Bruno Lemaitre, Toshiyuki Yamaji, Kentaro Hanada, John E. Burke, Ana Marija Jakšić, Brian D. McCabe, Paolo De Los Rios, Thorsten Hornemann, Giovanni D’Angelo, Vincenzo A. Gennarino

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Figure 6

CERT gain of function causes neurological defects in D. melanogaster.

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CERT gain of function causes neurological defects in D. melanogaster. (A...
(A) Schematic of transgenic dCERT flies on the w1118 background. dCERTWT, +WT; dCERTp.S149L, +SL; Chr, chromosome. (B) Quantification of endogenous or exogenous dCERT mRNA levels in flies on the w1118 background (Ctrl) in dCERT-transgenic flies. Data indicate the log2 fold change over Ctrl or +WT (n = 6; data are the mean ± SD). (C) Representative specimens of Ctrl, +WT, and +SL adult flies. Head length (HL), abdomen length (AL), and wing length (WL) were measured. Scale bars: 1 mm. (D) 3D rendering of a micro-CT scan of the heads from Ctrl, +WT, and +SL adults (frontal view). Brain volume is highlighted in pink. Body axes are dorsal (D) and left (L). Scale bars: 100 μm. (E) Z-projections of confocal stacks of whole-mount adult Ctrl, +WT, and +SL adult brains (frontal view) immunostained with anti-nc82. Scale bars: 100 μm. (F) HL/AL ratio of flies reared on DMSO or 10 μM HPA-12 (HPA) (Ctrl, n = 26 or 30, +WT n = 31 or 31, and +SL, n = 25 or 38). (G) Brain volume for Ctrl, +WT, and +SL flies as determined by confocal microscopy (n = 3–4; data are the mean ± SEM). (H) Mass spectrometry profile of sphingolipids in Ctrl, +WT, and +SL adult heads (n = 8). (I) Locomotion of Ctrl, +WT, and +SL flies plotted as total counts per 30 minutes over time (n = 16). (J) Locomotion of flies pretreated with 0 or 10 μM HPA-12 (n = 12). (K) Climbing ability of flies at 2, 9, 16, and 23 days post eclosion (dpe) after vigorous mechanical stress (n = 9). Data shown are the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA.