Second Generation γ-Secretase Modulators Exhibit Different Modulation of Notch β and Aβ Production

• Published in: The Journal of biological chemistry Vol. 287; no. 39; pp. 32640 - 32650
• Main Authors: Wanngren, Johanna, Ottervald, Jan, Parpal, Santiago, Portelius, Erik, Strömberg, Kia, Borgegård, Tomas, Klintenberg, Rebecka, Juréus, Anders, Blomqvist, Jenny, Blennow, Kaj, Zetterberg, Henrik, Lundkvist, Johan, Rosqvist, Susanne, Karlström, Helena
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21.09.2012; American Society for Biochemistry and Molecular Biology
• Subjects: Alzheimer Disease; Amyloid; Amyloid - genetics; Amyloid - metabolism; Amyloid beta-Protein Precursor; Amyloid beta-Protein Precursor - genetics; Amyloid beta-Protein Precursor - metabolism; Amyloid Precursor Protein; Amyloid Precursor Protein Secretases; Amyloid Precursor Protein Secretases - genetics; Amyloid Precursor Protein Secretases - metabolism; analogs & derivatives; Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; drug effects; Enzyme Processing; Female; Flurbiprofen; Flurbiprofen - pharmacology; Gamma-Secretase Modulators; genetics; HEK293 Cells; Humans; MALDI-TOF MS; Medicin och hälsovetenskap; metabolism; Mice; Molecular Bases of Disease; Neurodegenerative Diseases; Neurosciences; Neurovetenskaper; Non-Steroidal; Notch; pharmacology; Post-Translational; Protein Processing; Protein Processing, Post-Translational - drug effects; Protein Processing, Post-Translational - genetics; Protein Structure; Protein Structure, Tertiary; Receptors; Receptors, Notch - genetics; Receptors, Notch - metabolism; Sulindac; Sulindac - analogs & derivatives; Sulindac - pharmacology; Tertiary; Neurodegenerative Diseases; MALDI-TOF MS; Amyloid Precursor Protein; Notch; Gamma-Secretase Modulators; Alzheimer Disease; Enzyme Processing
• ISSN: 0021-9258; 1083-351X; 1083-351X
• DOI: 10.1074/jbc.M112.376541

The γ-secretase complex is an appealing drug target when the therapeutic strategy is to alter amyloid-β peptide (Aβ) aggregation in Alzheimer disease. γ-Secretase is directly involved in Aβ formation and determines the pathogenic potential of Aβ by generating the aggregation-prone Aβ42 peptide. Because γ-secretase mediates cleavage of many substrates involved in cell signaling, such as the Notch receptor, it is crucial to sustain these pathways while altering the Aβ secretion. A way of avoiding interference with the physiological function of γ-secretase is to use γ-secretase modulators (GSMs) instead of inhibitors of the enzyme. GSMs modify the Aβ formation from producing the amyloid-prone Aβ42 variant to shorter and less amyloidogenic Aβ species. The modes of action of GSMs are not fully understood, and even though the pharmacology of GSMs has been thoroughly studied regarding Aβ generation, knowledge is lacking about their effects on other substrates, such as Notch. Here, using immunoprecipitation followed by MALDI-TOF MS analysis, we found that two novel, second generation GSMs modulate both Notch β and Aβ production. Moreover, by correlating S3-specific Val-1744 cleavage of Notch intracellular domain (Notch intracellular domain) to total Notch intracellular domain levels using immunocytochemistry, we also demonstrated that Notch intracellular domain is not modulated by the compounds. Interestingly, two well characterized, nonsteroidal anti-inflammatory drugs (nonsteroidal anti-inflammatory drug), R-flurbiprofen and sulindac sulfide, affect only Aβ and not Notch β formation, indicating that second generation GSMs and nonsteroidal anti-inflammatory drug-based GSMs have different modes of action regarding Notch processing. Background: The γ-secretase complex is a drug target in the treatment of Alzheimer disease (AD). Results: Two novel second generation γ-secretase modulators (GSMs) modulate both Nβ and Aβ but not Notch intracellular domain (NICD) production. Conclusion: Second generation and NSAID-based GSMs have different modes of action regarding Notch processing. Significance: GSMs that do not affect NICD signaling are essential for the development of tolerable AD therapeutics.