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 1

CD163+ macrophages are associated with reduced vascular calcification in human carotid artery advanced atheromas.

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CD163+ macrophages are associated with reduced vascular calcification in...
(A–C) FCP sections from 82-year-old male. Low-power H&E image (A). High-power AR (B) and CD163 immunostaining (C) from the corresponding black rectangle in A. No CD163+ cells were observed around calcification stained as red in AR. (D–F) PR sections from 46-year-old male. Low-power H&E image (D). High-power images of AR (E) and CD163 immunostaining (F) from the corresponding black rectangle in D. Multiple CD163+ cells were observed around hemorrhagic area without any AR-positive lesions. (G and H) Seventy atheroma sections from 32 patients were classified into FCP or FA-Ca (n = 36, blue), FA or TCFA (n = 13, green), and PR or HR (n = 21, red), with the corresponding percentage of calcification (G) and CD163+ macrophages (H) per plaque area. (I–R) FA sections from 62-year-old male. Low-power Movat image (I). Mid-power image of H&E (J) of the corresponding black rectangle in I. High-power images of VK (K and O), AR (L and P), as well as immunostaining for CD68 (a general macrophage marker) (M and Q) and CD163 (N and R) of the corresponding red (K–N) and blue rectangles (O–R) in J. Note the absence of calcification in areas of CD163+ macrophages but its presence in CD163– areas. (S and T) CD163+CD68+ and CD163–CD68+ areas were determined in 70 sections (the method of outlining was in Supplemental Figure 1). Each CD163+CD68+ and CD163–CD68+ area was put in adjacent AR sections. Then AR-positive microcalcification areas in each field were digitally analyzed. The total area of CD163+CD68+ and CD163–CD68+ was similar (S); however, % microcalcification in CD163–CD68+ was greater than in CD163+CD68+ (T). Results are presented as median and interquartile range (G, H, S, and T). Kruskal-Wallis test followed by post hoc Dunn’s test (G and H) or Wilcoxon’s matched-pair signed-rank test (S and T) was applied. Data normality was tested by Shapiro-Wilk test. Scale bars: 2 mm (A and D), 0.2 mm (B, C, E, and F) 1 mm (I), 0.2 mm (J), 0.1 mm (K–R). AR, Alizarin Red; Ca, calcification; FA, fibroatheroma; FA-Ca, fibroatheroma with calcification; FCP, fibrocalcific plaque; HR, healed plaque rupture; PR, plaque rupture; TCFA, thin-cap fibroatheroma; VK, von Kossa.