CERT1 mutations perturb human development by disrupting sphingolipid homeostasis

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

CERT1 mutations perturb human development by disrupting sphingolipid homeostasis

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.This work was supported by the National Institute of Neurological Disorders and Stroke (NINDS), NIH (R01NS109858, to VAG); the Paul A. Marks Scholar Program at the Columbia University Vagelos College of Physicians and Surgeons (to VAG); a TIGER grant from the TAUB Institute at the Columbia Vagelos College of Physicians and Scientists (to VAG); the Swiss National Science Foundation (SNF 31003A-179371, to TH); the European Joint Program on Rare Diseases (EJP RD+SNF 32ER30-187505, to TH); the Swiss Cancer League (KFS-4999-02-2020, to GD); the EPFL institutional fund (to GD); the Kristian Gerhard Jebsen Foundation (to GD); the Swiss National Science Foundation (SNSF) (310030_184926, to GD); the Swiss Foundation for Research on Muscle Disease (FSRMM, to MAL); the Natural Science and Engineering Research Council of Canada (Discovery Grant 2020-04241, to JEB); the Italian Ministry of Health Young Investigator Grant (GR-2011-02347754, to EL); the Fondazione Istituto di Ricerca Pediatrica – Città della Speranza (18-04, to EL); the Wroclaw Medical University (SUB.E160.21.004, to RS); the National Science Centre, Poland (2017/27/B/NZ5/0222, to RS); Telethon Undiagnosed Diseases Program (TUDP) (GSP15001); the Temple Street Foundation/Children’s Health Foundation Ireland (RPAC 19-02, to IK); the Deutsche Forschungsgemeinschaft (DFG) (PO2366/2–1, to BP); the Instituto de Salud Carlos III, Spain (to ELM, EBS, and BMD); the National Natural Science Foundation of China (81871079 and 81730036, to HG and KX); and the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK), NIH (R01 DK115574, to SSC).The DEFIDIAG study is funded by grants from the French Ministry of Health in the framewok of the national French initiative for genomic medicine. The funders were not involved in the study design, data acquisition, analysis, or writing of the manuscript. Funding for the DECIPHER project was provided by Wellcome. The DDD study presents independent research commissioned by the Health Innovation Challenge Fund (grant number HICF-1009-003), a parallel funding partnership between Wellcome and the Department of Health, and the Wellcome Sanger Institute (grant number WT098051). The views expressed in this publication are those of the author(s) and not necessarily those of Wellcome or the Department of Health. The study has UK Research Ethics Committee approval (10/H0305/83, granted by the Cambridge South REC, and GEN/284/12, granted by the Republic of Ireland REC). The research team acknowledges the support of the National Institute for Health Research, through the Comprehensive Clinical Research Network.S

Flexible Instruction-Set Semantics via Abstract Monads (Experience Report)

Instruction sets, from families like x86 and ARM, are at the center of many ambitious formal-methods projects. Many verification, synthesis, programming, and debugging tools rely on formal semantics of instruction sets, but different tools can use semantics in rather different ways. The best-known work applying single semantics across diverse tools relies on domain-specific languages like Sail, where the language and its translation tools are specialized to the realm of instruction sets. In the context of the open RISC-V instruction-set family, we decided to explore a different approach, with semantics written in a carefully chosen subset of Haskell. This style does not depend on any new language translators, relying instead on parameterization of semantics over type-class instances. We have used a single core semantics to support testing, interactive proof, and model checking of both software and hardware, demonstrating that monads and the ability to abstract over them using type classes can support pleasant prototyping of ISA semantics

Effect of MacroRAFT Copolymer Adsorption on the Colloidal Stability of Layered Double Hydroxide Nanoparticles.

International audienceThe colloidal behavior of layered double hydroxide nanoparticles containing Mg2+ and Al3+ ions as intralayer cations and nitrates as counterions (MgAl-NO3-LDH) was studied in the presence of a short statistical copolymer of acrylic acid (AA) and butyl acrylate (BA) terminated with 4-cyano-4-thiothiopropylsulfanyl pentanoic acid (CTPPA) (P(AA7.5-stat-BA7.5)-CTPPA) synthesized by reversible addition–fragmentation chain-transfer (RAFT) polymerization. Surface charge properties and aggregation of the particles were investigated by electrophoresis and dynamic light scattering (DLS), respectively. The negatively charged P(AA7.5-stat-BA7.5)-CTPPA adsorbed strongly on the oppositely charged particles, leading to charge neutralization at the isoelectric point (IEP) and charge reversal at higher copolymer concentrations. The dispersions were unstable, i.e., fast aggregation of the MgAl-NO3-LDH occurred near the IEP while high stability was achieved at higher P(AA7.5-stat-BA7.5)-CTPPA concentrations. Atomic force (AFM) and transmission electron (TEM) microscopy imaging revealed that the platelets preferentially adopted a face-to-face orientation in the aggregates. While the stability of the bare particles was very sensitive to ionic strength, the P(AA7.5-stat-BA7.5)-CTPPA copolymer-coated particles were extremely stable even at high salt levels. Accordingly, the limited colloidal stability of bare MgAl-NO3-LDH dispersions was significantly improved by adding an appropriate amount of P(AA7.5-stat-BA7.5)-CTPPA to the suspension

Tailored microstructure and mechanical properties of nanocomposite films made from polyacrylic/LDH hybrid latexes synthesized by RAFT-mediated emulsion polymerization

International audienceLayered Double Hydroxide (LDH)-filled nanocomposites were processed from film-forming latexes synthesized by macroRAFT-assisted encapsulating emulsion polymerization (REEP). The microstructure and thermomechanical behavior of the polymer matrices and corresponding nanocomposites were investigated by TEM, FIB-SEM, SAXS and DMA. Strong ionic interactions created between acrylic acid groups induce lamellar nanostructuration of the P(AA-BA)-b-P(MA-BA) diblock copolymer matrix and are responsible for a high Young's modulus in the rubbery state that increases with the length (i.e., the molar mass) of the P(AA-BA) blocks. When filled with 16 wt% of LDH, the intrinsic structure of the matrix is lost and the mechanical behavior of the nanocomposites is solely driven by the LDH dispersion and ionic interactions. Two types of percolation (i.e., mesh size of the LDH network and mechanical reinforcement) have been achieved by playing with the degree of segregation of the LDH nanoplatelets within the material

Nanocomposite latexes containing layered double hydroxides via RAFT-assisted encapsulating emulsion polymerization

International audienceNanocomposite latex particles containing layered double hydroxide (LDH) platelets were synthesized using reversible addition–fragmentation chain transfer (RAFT) seeded emulsion polymerization. A random copolymer of acrylic acid (AA) and n-butyl acrylate (BA) was first synthesized by RAFT polymerization, and then electrostatically adsorbed on both nitrate- and carbonate-intercalated Mg2Al LDH particles to provide both colloidal stability and reactivatable groups from which the subsequent emulsion polymerization could proceed. The nitrate-intercalated LDH showed higher adsorption capacity than its carbonate counterpart because interlayer nitrate ions (in addition to those on the outer surface) were also displaced by the macroRAFT agent. The two macroRAFT agent-modified LDHs were then engaged in the emulsion polymerization of a hydrophobic monomer mixture (methyl acrylate (MA) and BA, 80/20 wt/wt) to form an encapsulating polymer shell. Cryogenic-transmission electron microscopy (cryo-TEM) showed successful encapsulation of the LDH nanoplatelets in the core of the latex particles, with the use of a hydrolytically-stable cationic initiator proving key to achieving high monomer conversions

Controlling the Morphology of Film-Forming, Nanocomposite Latexes Containing Layered Double Hydroxide by RAFT-Mediated Emulsion Polymerization

International audienceStable nanocomposite latexes with sandwich, encapsulated, or armored morphology were produced by starved-feed emulsion polymerization in the presence of layered double hydroxide (LDH) nanoparticles. Four statistical copolymers of acrylic acid (AA) and n-butyl acrylate (BA) were adsorbed on the LDH surface, and a film-forming methyl acrylate (MA)/BA (8:2 mass/mass) monomer feed was chosen to facilitate subsequent film formation under ambient conditions. P(AA17.5-co-BA17.5)-R (R = RAFT function) produced the sandwich morphology, while P(AA7.5-co-BA7.5)-R gave the encapsulated morphology, which is remarkable given that nonencapsulated morphologies are normally considered to be the preferred equilibrium structures for nanocomposite particles. Adsorption behavior and monomer conversion profiles were very similar for the two systems, and we tentatively ascribe the morphological difference to the higher density of RAFT functions in the P(AA7.5-co-BA7.5)-R system. A RAFT-free analogue produced armored latexes, highlighting the importance of the RAFT function for promoting growth of the polymer shell from the nanoparticle surface

Multicolour Optical Coding from a Series of Luminescent Lanthanide Complexes with a Unique Antenna

International audienceThe bis‐tetrazolate‐pyridine ligand $H_{2}$pytz sensitises efficiently the visible and/or near‐IR luminescence emission of ten lanthanide cations (Pr, Nd, Sm, Eu, Tb, Dy, Ho, Er, Tm, Yb). The $Ln^{III}$ complexes present sizeable quantum yields in both domains with a single excitation source. The wide range of possible colour combinations in water, organic solvents and the solid state makes the complexes very attractive for labelling and encoding