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 4

BACE2-dependent changes in VEGFR3 signaling in LECs.

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BACE2-dependent changes in VEGFR3 signaling in LECs. (A) Schematic for V...
(A) Schematic for VEGFR3 signaling. Upon ligand binding, VEGFR3 dimerizes, resulting in intracellular autophosphorylation and activation of the downstream genes FOXC2 and DLL4. V, verubecestat, inhibitor of BACE2. Gene expression levels of (B) DLL4 and FOXC2 and (C) VEGFR3 after the application of DMSO, 100 nM verubecestat (V), and VEGF-C. (D and E) Gene expression levels of DLL4, FOXC2, VEGFR3, BACE1, and BACE2 after BACE knockdown (siB1/siB2) without or with (+V) subsequent verubecestat application. All dot plots were normalized on the control mean and depict mean and SD alongside the P values calculated by unpaired t tests against the DMSO control (B and C) or by 1-way ANOVA (D and E), in both cases followed by Bonferroni’s multiple-comparison test. *P < 0.05; **P < 0.01; ****P < 0.0001. P values are only indicated where significance was observed. In B, 1 data point was excluded from the DLL4 expression/VEGF-C156S data set, since it was identified as an outlier via the ROUT method. Data are derived from n = 6 (B and C) or n = 4 (D and E) biological replicates.