Bone-derived PDGF-BB drives brain vascular calcification in male mice
Jiekang Wang, … , Xu Cao, Mei Wan
Jiekang Wang, … , Xu Cao, Mei Wan
Published October 10, 2023
Citation Information: J Clin Invest. 2023;133(23):e168447. https://doi.org/10.1172/JCI168447.
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Research Article Bone biology Vascular biology

Bone-derived PDGF-BB drives brain vascular calcification in male mice

Abstract

Brain vascular calcification is a prevalent age-related condition often accompanying neurodegenerative and neuroinflammatory diseases. The pathogenesis of large-vessel calcifications in peripheral tissue is well studied, but microvascular calcification in the brain remains poorly understood. Here, we report that elevated platelet-derived growth factor BB (PDGF-BB) from bone preosteoclasts contributed to cerebrovascular calcification in male mice. Aged male mice had higher serum PDGF-BB levels and a higher incidence of brain calcification compared with young mice, mainly in the thalamus. Transgenic mice with preosteoclast-specific Pdgfb overexpression exhibited elevated serum PDGF-BB levels and recapitulated age-associated thalamic calcification. Conversely, mice with preosteoclast-specific Pdgfb deletion displayed diminished age-associated thalamic calcification. In an ex vivo cerebral microvascular culture system, PDGF-BB dose-dependently promoted vascular calcification. Analysis of osteogenic gene array and single-cell RNA-Seq (scRNA-Seq) revealed that PDGF-BB upregulated multiple osteogenic differentiation genes and the phosphate transporter Slc20a1 in cerebral microvessels. Mechanistically, PDGF-BB stimulated the phosphorylation of its receptor PDGFRβ (p-PDGFRβ) and ERK (p-ERK), leading to the activation of RUNX2. This activation, in turn, induced the transcription of osteoblast differentiation genes in PCs and upregulated Slc20a1 in astrocytes. Thus, bone-derived PDGF-BB induced brain vascular calcification by activating the p-PDGFRβ/p-ERK/RUNX2 signaling cascade in cerebrovascular cells.

Authors

Jiekang Wang, Ching-Lien Fang, Kathleen Noller, Zhiliang Wei, Guanqiao Liu, Ke Shen, Kangping Song, Xu Cao, Mei Wan

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Figure 1

Aged male mice develop brain calcification at the thalamic regions.

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Aged male mice develop brain calcification at the thalamic regions. (A) ...
(A) Axial and coronal SWI sequence images of brains from 3-month-old male mice (3M-M), 22-month-old male mice (22M-M), 3-month-old female mice (3M-F), and 22-month-old female mice (22M-F). Calcifications (yellow arrows) were observed as black structures in the thalamic region on susceptibility-weighted images. n = 6–9. (B) Calcification incidence calculation based on the SWI analysis in A. (C) The volume of calcification in different brain regions, including the cortex, hippocampus, thalamus, and hypothalamus, in 22-month-old male mice was determined on the basis of SWI analysis. n = 5. (D) Quantification of the volume of calcification in individual serial sections (750 μm thick) by SWI throughout the entire thalamus, from caudal to rostral aspects. n = 5. (E) Alizarin red staining of brain tissue sections from 3- and 22-month-old male mice. Boxed areas are shown at higher magnification (×10) in the corresponding panels on the right. The calcification nodule is shown in red. n = 9. (F) Calculation of the calcification incidence in 3- and 22-month-old male mice based on the histology stainings. n = 9. (G) Double-immunofluorescence staining of frozen brain tissue sections from 3- and 22-month-old male mice using antibodies against CD13 and OPN. n = 5. (H) Quantification of the volume of the OPN+ calcified nodules in G. n = 5. The Imaris 3D reconstruction method was used to quantify the number and volume of calcification. (I and J) Double-immunofluorescence staining of frozen brain tissue sections from 3- and 22-month-old male mice using antibodies against OPN and OCN (I) or OPN and CatK (J). n = 3. Scale bars: 100 μm (G, I, and J). (K) Immunofluorescence staining was performed on brain sections from male mice at 3 and 22 months of age using an antibody against ALPL. n = 3. Scale bar: 100 μm. Data are shown as the mean ± SD. **P < 0.01, by unpaired, 2-tailed Student’s t test (H).