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

Authors

Simone M. Crivelli, University of KentuckyFollow
Qian Luo, Maastricht University, the Netherlands
Jo A. A. Stevens, Maastricht University, the Netherlands
Caterina Giovagnoni, Maastricht University, the Netherlands
Daan van Kruining, Maastricht University, the Netherlands
Gerard Bode, Maastricht University, the Netherlands
Sandra den Hoedt, Erasmus MC University, the Netherlands
Barbara Hobo, Netherlands institute for Neuroscience, the Netherlands
Anna-Lena Scheithauer, University of Bonn, Germany
Jochen Walter, University of Bonn, Germany
Monique T. Mulder, Erasmus MC University, the Netherlands
Christopher Exley, Keele University, UK
Matthew Mold, Keele University, UK
Michelle M. Mielke, Mayo Clinic Rochester
Helga E. De Vries, Amsterdam UMC, the Netherlands
Kristiaan Wouters, Maastricht University, the Netherlands
Daniel L. A. van den Hove, University of Wuerzburg, Germany
Dusan Berkes, Slovak University of Technology, Slovak Republic
María Dolores Ledesma, Centro de Biología Molecular Severo Ochoa, Spain
Joost Verhaagen, Netherlands institute for Neuroscience, the Netherlands
Mario Losen, Maastricht University, the Netherlands
Erhard Bieberich, University of KentuckyFollow
Pilar Martinez-Martinez, Maastricht University, the Netherlands

Abstract

BACKGROUND: Dysregulation 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 which are 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: A plasmid expressing CERT_L, the long isoform of CERTs, was used to study the interaction of CERT_L with amyloid precursor protein (APP) by co-immunoprecipitation and immunofluorescence in HEK cells. The recombinant CERT_L protein was employed to study interaction of CERT_L with amyloid-β (Aβ), Aβ aggregation process in presence of CERT_L, and the resulting changes in Aβ toxicity in neuroblastoma cells. CERT_L was overexpressed in neurons by adeno-associated virus (AAV) in a mouse model of familial AD (5xFAD). Ten weeks after transduction, animals were challenged with behavior tests for memory, anxiety, and locomotion. At week 12, brains were investigated for sphingolipid levels by mass spectrometry, plaques, and neuroinflammation by immunohistochemistry, gene expression, and/or immunoassay.

RESULTS: Here, we report that CERT_L binds to APP, modifies Aβ aggregation, and reduces Aβ neurotoxicity in vitro. Furthermore, we show that intracortical injection of AAV, mediating the expression of CERT_L, decreases levels of ceramide d18:1/16:0 and increases sphingomyelin levels in the brain of male 5xFAD mice. CERT_L in vivo over-expression has a mild effect on animal locomotion, decreases Aβ formation, and modulates microglia by decreasing their pro-inflammatory phenotype.

CONCLUSION: Our results demonstrate a crucial role of CERT_L 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.

Document Type

Article

Publication Date

2-17-2021

Notes/Citation Information

Published in Alzheimer's Research & Therapy, v. 13, issue 1, article no. 45.

© The Author(s) 2021

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Digital Object Identifier (DOI)

https://doi.org/10.1186/s13195-021-00780-0

Funding Information

This work was supported by grants to NMdW, SdH, MTM, JW, AR, PMM, JV, and HEV from ZonMw Memorabel program (projectnr: 733050105). PMM is also supported by the International Foundation for Alzheimer Research (ISAO) (projectnr: 14545). SMC received a travel grant support from Alzheimer Nederlands foundation (AN) to spend a month in the laboratory of EB, University of Kentucky, Lexington KY, USA. Aspects of this work were supported by the grants NIH R01AG034389, R01NS095215, and R56AG064234; VA I01BX003643 to EB. MMM was supported by R01AG049704.

Related Content

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Repository Citation

Crivelli, Simone M.; Luo, Qian; Stevens, Jo A. A.; Giovagnoni, Caterina; van Kruining, Daan; Bode, Gerard; den Hoedt, Sandra; Hobo, Barbara; Scheithauer, Anna-Lena; Walter, Jochen; Mulder, Monique T.; Exley, Christopher; Mold, Matthew; Mielke, Michelle M.; De Vries, Helga E.; Wouters, Kristiaan; van den Hove, Daniel L. A.; Berkes, Dusan; Ledesma, María Dolores; Verhaagen, Joost; Losen, Mario; Bieberich, Erhard; and Martinez-Martinez, Pilar, "CERT_L Reduces C16 Ceramide, Amyloid-β Levels, and Inflammation in a Model of Alzheimer’s Disease" (2021). Physiology Faculty Publications. 169.
https://uknowledge.uky.edu/physiology_facpub/169