Carbonic anhydrases II and XII are up-regulated in osteoclast-like cells in advanced human atherosclerotic plaques—Tampere Vascular Study

Figures & data

Table I. Genome-wide gene expression analysis of genes related to bone formation in arterial plaques relative to non-atherosclerotic internal thoracic artery control samples in Tampere Vascular Study.

	Fold-change
Gene	Carotid artery	Femoral artery	Aorta
Osteoclastogenesis:
c-fms	3.3***	2.8***	3.0***
RANK	−1.0	1.0	1.0
RANKL2	1.1*	1.1	1.1
Osteoprotegerin	4.8***	1.7	2.3***
Osteonectin	1.2	1.6*	1.2*
Osteopontin	64.8***	42.6***	41.6***
MMP9	29.5***	21.7***	19 9***
Cathepsin K	2.2**	2.8**	1.5
ACP5	20.4***	15.3***	8.1**
CLCN7	2.5***	2.1***	2.1***
TCIRG1_1	1.2***	1.2*	1.2*
OSTM1	1.3***	1.1	1.1
Osteoblastogenesis:
RUNX2	–	–	−1.1
TWIST1	–	−1.1	−1.1
WNT1	1.1	–	–
Dlx5	1.1*	1.1*	–
SP7	1.1	1.1	–
FOSL1	–	1.1	1.1

RANK=Homo sapiens tumor necrosis factor receptor superfamily, member 11a, activator of NF-κB (TNFRSF11A), mRNA; RANKL2 = Homo sapiens tumor necrosis factor (ligand) superfamily, member 11 (TNFSF11), transcript variant 2, mRNA; ACP5 = Homo sapiens acid phosphatase 5, tartrate-resistant (ACP5), mRNA; MMP9 = Homo sapiens matrix metalloproteinase 9 (gelatinase B, 92 κDa gelatinase, 92 kDa type IV collagenase) (MMP9), mRNA; OSTM1=Homo sapiens osteopetrosis-associated transmembrane protein 1 (OSTM1), mRNA; CLCN7 = Homo sapiens chloride channel 7 (CLCN7), mRNA; TCIRG1_1=Homo sapiens T-cell, immune regulator 1, ATPase, H+ transporting, lysosomal V0 protein a isoform 3 (TCIRG1), transcript variant 1, mRNA; c-fms = Homo sapiens colony-stimulating factor 1 receptor, formerly McDonough feline sarcoma viral (v-fms) oncogene homolog (CSF1R), mRNA; RUNX2 = runt-related transcription factor 2; TWIST = twist homolog 1; WNT = wingless-type MMTV integration site family, member 1; Dlx5 = distal-less homeobox 5; SP7 = Sp7 transcription factor; FOSL1 =FOS-like antigen 1. *P < 0.05, *P< 0.01, ***P< 0.0001 relative to control, Mann-Whitney U-test.

Figure 1. CA2 and CA12 expression in control internal thoracic artery (ITA), carotid, femoral, and aortic plaque samples. Gene expression value is the normalized average gene intensity for each group measured in the Illumina Expression BeadChips. *P < 0.05, **P < 0.01, ***P < 0.001 relative to control (ITA), Mann-Whitney U-test.

Table II. Genome-wide gene expression analysis of carbonic anhydrases in arterial plaques relative to non-atherosclerotic internal thoracic artery control samples in Tampere Vascular Study.

	Fold-change
Gene	Carotid artery	Femoral artery	Aorta
CA1	−1.3	−1.3	−1.4**
CA2	2.3***	2.2**	1.9**
CA3	−1.2	−1.2	−1.1
CA4	−1.2*	−1.1	1.1
CA5A	1.0	1.1	1.0
CA5B	1.1	−1.2	−1.2
CA6	1.1	−1.2	1.1
CA7	−1.1	1.1	−1.1
CAS	1.0	−1.1	1.0
CA9	1.1	−1.0	1.0
CA10	1.0	−1.0	1.1
CA11	1.0	1.0	−1.0
CA12	3.4***	3.7***	2.7***
CA13	−1.0	1.1	1.0
CAM	−1.0	−1.1	−1.0

* P< 0.05, *P< 0.01, *** P< 0.0001 relative to control, Mann-Whitney U-test.

Figure 2. CA2 and CA12 expression in atherosclerotic plaque and control internal thoracic artery (ITA) samples measured with TaqMan Low-Density Array (LDA). *P < 0.05, **P < 0.01, ***P < 0.001 relative to control (ITA), Mann-Whitney U-test.

Figure 3. Expression of CA2 and CA12 in control internal thoracic artery (ITA), carotid, aortic, and femoral plaque samples measured with TaqMan Low-Density Array (LDA). *P < 0.05, **P < 0.01, ***P < 0.001 relative to control (ITA), Mann-Whitney U-test.

Figure 4. Expression of CAII and CAXII proteins in human atherosclerotic plaques in human aortic (A), femoral (B), and carotid (C) plaques. Samples from internal thoracic artery served as controls (D). Arrows indicate typical positively stained cells. The stage of atherosclerosis was classified according to American Heart Association (AHA) classification (type I–VI). 100× magnification.

Figure 5. Double immunofluorescence and mirror image section images demonstrating co-localization of CAII and CAXII proteins with CD68 in human atherosclerotic plaques in representative carotid plaques. CAII labeling is cytosolic while CAXII labeling is strong in the cell membrane.

Figure 6. Mirror image section images demonstrating co-localization of CAXII protein with TRAP in human atherosclerotic plaques in representative carotid plaques. Corresponding cells, cell clusters, and microanatomic structures are indicated by an arrow-head, numbers 1–4, stars, and an asterisk.