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.
View: Text | PDF
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

×

Figure 2

Preosteoclast-secreted PDGF-BB is sufficient to induce brain calcification.

Options: View larger image (or click on image) Download as PowerPoint
Preosteoclast-secreted PDGF-BB is sufficient to induce brain calcificati...
(A–C) ELISA measurements of serum PDGF-BB concentrations in 3- and 22-month-old male mice (n = 5) (A), 3- and 22-month-old female mice (n = 6) (B), and 6-month-old PdgfbcTG mice and WT littermates (n = 4) (C). (D) Calculation of calcification incidence in 6-month-old PdgfbcTG mice and WT littermates. n = 9. (E) Representative images of Alizarin red staining of brain tissue sections from 6-month-old PdgfbcTG mice and WT littermates. The calcification nodule is shown in red. Boxed areas are shown at a higher magnification (×10) in the corresponding panels on the right. n = 9. (F) Double-immunofluorescence staining of frozen brain tissue sections from 6-month-old PdgfbcTG using antibodies against CD13 and OPN. n = 3. (G) OPN expression in hippocampus, thalamus, and cortex from PdgfbcTG mice and WT littermates was measured by Western blot analysis. n = 5. (H) The relative intensity from G was calculated using ImageJ. n = 5. (I) Immunofluorescence staining of frozen brain tissue sections from PdgfbcTG mice and WT littermates using antibodies against ALPL. n = 3. Boxed areas are shown at higher magnification (×5) in the corresponding panels on the right. n = 3. Scale bars: 100 μm (E and I); 200 μm (F). Data are shown as the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001, by unpaired, 2-tailed Student’s t test (A–C and H).