Ceramide Transporter CERT Is Involved in Muscle Insulin Signaling Defects Under Lipotoxic Conditions

Bandet, Cécile; Mahfouz, Rana; Véret, Julien; Sotiropoulos, Athanassia; Poirier, Maxime; Giussani, Paola; Campana, Mélanie; Philippe, Erwann; Błachnio-Zabielska, Agnieszka; Ballaire, Raphaëlle; Lièpvre, Xavier Le; Bourron, Olivier; Berkeš, Dušan; Górski, Jan; Ferré, Pascal; Stunff, Hervé Le; Foufelle, Fabienne; Hajduch, Éric

doi:10.2337/db17-0901

Cited by 37 publications

(49 citation statements)

References 55 publications

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“…In this study we report that increasingCer tra cking from the ER to the Golgi by overexpressing CERT L , reversed the pathological increaseof Cer in the cortex. A similar effect of CERTs overexpression in Cer elevation state has been reported in alipotoxic mousemodel where muscle cells in overload Cer statuswere rescued by increasing the expression of hCERT (52). In AD brains, we observed that CERT L reduced 11% of the total Cer content restoring it close to normal levels.…”

Section: Discussionsupporting

confidence: 87%

“…It has been consistently reported that 5xFAD microglia are polarized towards a more pro-in ammatory status, in response to the extensive plaque formation. Consequently, the Iba1 microglia marker is highly expressed in AD models [52,53]. Our ndings suggest thatCERT L could play a role in the cross-talk between neurons and microglia.…”

Section: Discussionmentioning

confidence: 66%

“…AAV-CERT L decreased Cer d18:1/16:0and increased SM levels in the cortex.One of the main functions of CERTs is to shuttle Cerfrom the ER to the Golgi (22). It has been reported that toxic increase ofCer species in the muscle can be attenuated by overexpressing hCERTcDNA (52).HPLC-MS/MS analysis of brain cortex tissuerevealed a signi cant reduction of Cerd18:1/16:0 level due to CERT L overexpression (p<0.05).…”

Section: Resultsmentioning

confidence: 96%

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CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

Crivelli

Luo

Stevens

et al. 2020

Preprint

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Background: Deregulation of ceramide and sphingomyelinlevels have been suggested tocontribute tothe pathogenesis of Alzheimer’s disease (AD).Ceramide transfer proteins (CERTs) are ceramide carriers, crucial for ceramide and sphingomyelin balance in cells.Extracellular forms of CERTs co-localize with amyloid-β (Aβ) plaques in AD brains. To date, the significance of these observations for the pathophysiology of AD remains uncertain.Methods: The plasmid expressing CERTL, the long isoform of CERTs, was used to study the interaction of CERTLwith amyloid precursor protein (APP) by co-immunoprecipitation and immunofluorescencein HEK cells.The recombinant CERTL protein wasemployed to study interaction of CERTLwith amyloid-β (Aβ), Aβ aggregation process in presence of CERTL, and the resulting changes inAβ toxicity in neuroblastoma cells. CERTLwas overexpressed in neurons by adeno associatedvirus (AAV) in a familial mouse model of familial AD (5xFAD). Ten weeks after transduction animal were challenged with behavior tests for memory, anxiety and locomotion. At week twelve brains were investigated for sphingolipid levels by mass spectrometry, plaques and neuroinflammation by immunohistochemistry, gene expression and/or immunoassay.Results:Here, we report that CERTL, binds to APP, modifies Aβ aggregation and reduces Aβ neurotoxicity in vitro. Furthermore, we show that intracortical injection of AAV, mediating the expression of CERTL, decreases levels of ceramide d18:1/16:0 and increases sphingomyelin levels in the brain of male transgenic mice, modelling familial AD (5xFAD). CERTLin vivo over-expression hasa mild effect on animal locomotion and decreases Aβ formation and modulates microglia by decreasing their pro-inflammatory phenotype.Conclusion: Our results demonstratea crucial role of CERTL in regulatingceramidelevels in the brain, in amyloid plaque formation and neuroinflammation,thereby opening research avenuesfor therapeutic targets of AD and other neurodegenerative diseases.

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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 66%

Section: Resultsmentioning

confidence: 96%

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CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

Crivelli

Luo

Stevens

et al. 2020

Preprint

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“…Beyond its primary function to deliver ceramide from the ER to the Golgi apparatus, CERT participates in various biological events, including polyploid cancer cell death 32 , EGF receptor signaling 33 , lipotoxicity and glucolipotoxicity in islet β-cells 34,35 , muscle insulin signaling 36 , stress-induced Golgi disassembly 37 , protein secretions 38 , phosphoinositide turnover at the trans Golgi network 39 , cytotoxic autophagy 40 , and senescence 41 . Additionally, pharmacological or genetic inhibition of CERT negatively affects the proliferation of several types of intracellular pathogens [42][43][44][45][46] .…”

Section: Discussionmentioning

confidence: 99%

Natural ligand-nonmimetic inhibitors of the lipid-transfer protein CERT

et al. 2019

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Lipid transfer proteins mediate inter-organelle transport of membrane lipids at organelle contact sites in cells, playing fundamental roles in the lipidome and membrane biogenesis in eukaryotes. We previously developed a ceramide-mimetic compound as a potent inhibitor of the ceramide transport protein CERT. Here we develop CERT inhibitors with structures unrelated to ceramide. To this aim, we identify a seed compound with no ceramide-like structure but with the capability of forming a hydrogen-bonding network in the ceramidebinding START domain, by virtual screening of~3 × 10 6 compounds. We also establish a surface plasmon resonance-based system to directly determine the affinity of compounds for the START domain. Then, we subject the seed compound to a series of in silico docking simulations, efficient chemical synthesis, affinity analysis, protein-ligand co-crystallography, and various in vivo assays. This strategy allows us to obtain ceramide-unrelated compounds that potently inhibited the function of CERT in human cultured cells.

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“…One of the main functions of CERTs is to shuttle Cer from the ER to the Golgi (22). It has been reported that toxic increase of Cer species in the muscle can be attenuated by overexpressing hCERT cDNA (52). HPLC-MS/MS analysis of brain cortex tissue revealed a signi cant reduction of Cer d18:1/16:0 level due to CERT L overexpression (p < 0.05).…”

Section: Resultsmentioning

confidence: 97%

CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

Crivelli

Luo

Stevens

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Deregulation of ceramide and sphingomyelin levels have been suggested to contribute to the pathogenesis of Alzheimer’s disease (AD). Ceramide transfer proteins (CERTs) are ceramide carriers, crucial for ceramide and sphingomyelin balance in cells. Extracellular forms of CERTs co-localize with amyloid-β (Aβ) plaques in AD brains. To date, the significance of these observations for the pathophysiology of AD remains uncertain.Methods: The plasmid expressing CERTL, the long isoform of CERTs, was used to study the interaction of CERTL with amyloid precursor protein (APP) by co-immunoprecipitation and immunofluorescence in HEK cells. The recombinant CERTL protein was employed to study interaction of CERTL with amyloid-β (Aβ), Aβ aggregation process in presence of CERTL, and the resulting changes in Aβ toxicity in neuroblastoma cells. CERTL was overexpressed in neurons by adeno associated virus (AAV) in a familial mouse model of familial AD (5xFAD). Ten weeks after transduction animal were challenged with behavior tests for memory, anxiety and locomotion. At week twelve brains were investigated for sphingolipid levels by mass spectrometry, plaques and neuroinflammation by immunohistochemistry, gene expression and/or immunoassay.Results: Here, we report that CERTL, binds to APP, modifies Aβ aggregation and reduces Aβ neurotoxicity in vitro. Furthermore, we show that intracortical injection of AAV, mediating the expression of CERTL, decreases levels of ceramide d18:1/16:0 and increases sphingomyelin levels in the brain of male transgenic mice, modelling familial AD (5xFAD). CERTL in vivo over-expression has a mild effect on animal locomotion and decreases Aβ formation and modulates microglia by decreasing their pro-inflammatory phenotype.Conclusion: Our results demonstrate a crucial role of CERTL in regulating ceramide levels in the brain, in amyloid plaque formation and neuroinflammation, thereby opening research avenues for therapeutic targets of AD and other neurodegenerative diseases.

show abstract

Ceramide Transporter CERT Is Involved in Muscle Insulin Signaling Defects Under Lipotoxic Conditions

Cited by 37 publications

References 55 publications

CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

Natural ligand-nonmimetic inhibitors of the lipid-transfer protein CERT

CERTL Reduces C16 Ceramide, Amyloid-β Levels and Inflammation in a Model of Alzheimer's Disease

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