CD163+ macrophages restrain vascular calcification, promoting the development of high-risk plaque
Atsushi Sakamoto, … , Renu Virmani, Aloke V. Finn
Atsushi Sakamoto, … , Renu Virmani, Aloke V. Finn
Published January 31, 2023
Citation Information: JCI Insight. 2023;8(5):e154922. https://doi.org/10.1172/jci.insight.154922.
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Research Article Cell biology Vascular biology

CD163+ macrophages restrain vascular calcification, promoting the development of high-risk plaque

Abstract

Vascular calcification (VC) is concomitant with atherosclerosis, yet it remains uncertain why rupture-prone high-risk plaques do not typically show extensive calcification. Intraplaque hemorrhage (IPH) deposits erythrocyte-derived cholesterol, enlarging the necrotic core and promoting high-risk plaque development. Pro-atherogenic CD163+ alternative macrophages engulf hemoglobin:haptoglobin (HH) complexes at IPH sites. However, their role in VC has never been examined to our knowledge. Here we show, in human arteries, the distribution of CD163+ macrophages correlated inversely with VC. In vitro experiments using vascular smooth muscle cells (VSMCs) cultured with HH-exposed human macrophage — M(Hb) — supernatant reduced calcification, while arteries from ApoE–/– CD163–/– mice showed greater VC. M(Hb) supernatant–exposed VSMCs showed activated NF-κB, while blocking NF-κB attenuated the anticalcific effect of M(Hb) on VSMCs. CD163+ macrophages altered VC through NF-κB–induced transcription of hyaluronan synthase (HAS), an enzyme that catalyzes the formation of the extracellular matrix glycosaminoglycan, hyaluronan, within VSMCs. M(Hb) supernatants enhanced HAS production in VSMCs, while knocking down HAS attenuated its anticalcific effect. NF-κB blockade in ApoE–/– mice reduced hyaluronan and increased VC. In human arteries, hyaluronan and HAS were increased in areas of CD163+ macrophage presence. Our findings highlight an important mechanism by which CD163+ macrophages inhibit VC through NF-κB–induced HAS augmentation and thus promote the high-risk plaque development.

Authors

Atsushi Sakamoto, Rika Kawakami, Masayuki Mori, Liang Guo, Ka Hyun Paek, Jose Verdezoto Mosquera, Anne Cornelissen, Saikat Kumar B. Ghosh, Kenji Kawai, Takao Konishi, Raquel Fernandez, Daniela T. Fuller, Weili Xu, Aimee E. Vozenilek, Yu Sato, Hiroyuki Jinnouchi, Sho Torii, Adam W. Turner, Hirokuni Akahori, Salome Kuntz, Craig C. Weinkauf, Parker J. Lee, Robert Kutys, Kathryn Harris, Alfred Lawrence Killey, Christina M. Mayhew, Matthew Ellis, Leah M. Weinstein, Neel V. Gadhoke, Roma Dhingra, Jeremy Ullman, Armella Dikongue, Maria E. Romero, Frank D. Kolodgie, Clint L. Miller, Renu Virmani, Aloke V. Finn

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Figure 9

CD163 activity and its association with NF-κB signaling, HA synthesis, and calcification in human atherosclerotic arteries.

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CD163 activity and its association with NF-κB signaling, HA synthesis, a...
(A–F) CTO section of left circumflex artery (LCX) from 37-year-old man who died from an acute coronary syndrome (culprit plaque rupture was found in Left anterior descending artery, LAD). Low-power Movat (A) and high-power H&E (B) image of the corresponding black rectangle in A. Yellow-dot border of occluded lumen with thrombus. Fibrin deposition and neo-angiogenesis (*) in the occluded lumen were observed. CD163 (red) (C and D) and HA (red)/HAS1 (green) (E and F) immunofluorescence images of the adjacent sections of A. (G–K) Representative immunoblotting (G) and summary of densitometry analysis (H–K) for CD163, p-p65 (Ser536), t-p65, HAS1, and β-actin of protein extracted from human carotid atheroma expressing high or low levels of CD163 (n = 6 per group). *P < 0.05, **P < 0.01. (L) X-ray image of postmortem heart from 41-year-old African American with WT allele of the rs7136716 (AA genotype). White arrowheads indicate severe calcification in the coronary tree. (M) X-ray image of postmortem heart from 44-year-old African American with 2 copies of the minor allele for rs7136716 (GG genotype) without visible coronary calcification. Pathology of coronary arteries revealed triple-vessel disease. (N) Summary of ex vivo x-ray–based calcification score in age-matched AA versus GG genotype carriers (n = 15 per group). (O–S) Representative histopathology images of coronary artery sections (H&E) from AA (O) and GG (P) genotype carriers. Percentage area stenosis (Q), total calcification area (R), and % calcification/plaque area (S) between 2 genotype carriers. Results are presented as the mean ± standard deviation (H, I, K, and Q) or median and interquartile range (J, N, R, and S). T test (H, I, K, and Q) or Mann-Whitney test (J, N, R, and S) was conducted for statistical analysis. Data normality was tested by Shapiro-Wilk test. Scale bars: 0.5 mm (A, C, and E), 0.1 mm (B, D, and F), 0.5 mm (O and P).