The Alzheimer’s disease–linked protease BACE2 cleaves VEGFR3 and modulates its signaling
Andree Schmidt, … , Bettina Schmid, Stefan F. Lichtenthaler
Andree Schmidt, … , Bettina Schmid, Stefan F. Lichtenthaler
Published June 18, 2024
Citation Information: J Clin Invest. 2024;134(16):e170550. https://doi.org/10.1172/JCI170550.
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Research Article Aging

The Alzheimer’s disease–linked protease BACE2 cleaves VEGFR3 and modulates its signaling

Abstract

The β-secretase β-site APP cleaving enzyme (BACE1) is a central drug target for Alzheimer’s disease. Clinically tested, BACE1-directed inhibitors also block the homologous protease BACE2. Yet little is known about physiological BACE2 substrates and functions in vivo. Here, we identify BACE2 as the protease shedding the lymphangiogenic vascular endothelial growth factor receptor 3 (VEGFR3). Inactivation of BACE2, but not BACE1, inhibited shedding of VEGFR3 from primary human lymphatic endothelial cells (LECs) and reduced release of the shed, soluble VEGFR3 (sVEGFR3) ectodomain into the blood of mice, nonhuman primates, and humans. Functionally, BACE2 inactivation increased full-length VEGFR3 and enhanced VEGFR3 signaling in LECs and also in vivo in zebrafish, where enhanced migration of LECs was observed. Thus, this study identifies BACE2 as a modulator of lymphangiogenic VEGFR3 signaling and demonstrates the utility of sVEGFR3 as a pharmacodynamic plasma marker for BACE2 activity in vivo, a prerequisite for developing BACE1-selective inhibitors for safer prevention of Alzheimer’s disease.

Authors

Andree Schmidt, Brian Hrupka, Frauke van Bebber, Sanjay Sunil Kumar, Xiao Feng, Sarah K. Tschirner, Marlene Aßfalg, Stephan A. Müller, Laura Sophie Hilger, Laura I. Hofmann, Martina Pigoni, Georg Jocher, Iryna Voytyuk, Emily L. Self, Mana Ito, Kana Hyakkoku, Akimasa Yoshimura, Naotaka Horiguchi, Regina Feederle, Bart De Strooper, Stefan Schulte-Merker, Eckhard Lammert, Dieder Moechars, Bettina Schmid, Stefan F. Lichtenthaler

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Figure 6

BACE2 inhibition reduces murine plasma sVEGFR3.

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BACE2 inhibition reduces murine plasma sVEGFR3. Volcano plots of proteom...
Volcano plots of proteomic analysis of murine plasma from (A) compound 89–treated (Cpd89) versus vehicle-treated (veh) mice and (B) LY2811376-treated versus vehicle-treated mice (n = 13, treated; n = 14, veh). VEGFR3 is highlighted in red. (C) Corresponding extracted LFQ intensities of sVEGFR3 and (D) MSD-assay quantifications of sVEGFR3. Plasma sVEGFR3 (E) and plasma sSEZ6L (F) levels in 8–10 B1KO, B2KO, and respective WT mice with (blue) or without (black) 3 days of 50 mg/kg per os twice a day verubecestat dosing. (G) Plasma levels of VEGFR3 and SEZ6L during 7 days of 0.1% dietary verubecestat (average drug intake, 97 mg/kg/d; n = 6 per group, all male, age: 7–10 weeks), respective to untreated control levels. Two-sided Student’s t tests with a permutation-based FDR correction (FDR < 0.05; indicated by hyperbolic curves) were used for volcano plots (A and B). Proteins with P < 0.05 are shown as red circles. (C) Significance after FDR correction is indicated with plus signs. All dot plots were normalized on the control mean and depict the SD alongside the calculated P values, calculated by 1-way (D and G) or 2-way (E and F) ANOVA with Bonferroni’s multiple-comparison test. *P < 0.05; ***P < 0.001; ****P < 0.0001. P values are only indicated where significance could be observed. Number of biological replicates in E and F was 9, except for Bace1-WT + verubecestat (n = 8) and for Bace2WT without verubecestat (n = 10).