Chronic kidney disease induces a distinct lipidomic signature and accelerates atherosclerosis progression in a novel minipig model

Figures

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  Fig. 1. Porcine CKD study: (A): experimental timeline and animal interventions. (B): schematic of the left kidney showing selective clamping of the cranial polar branch (crPB) and the anterior segmental branch of the caudal polar branch (CaPB) of the left renal artery (LRA). (C): laparoscopic port configuration for instrument triangulation. (D): intraoperative view of the CrPB clamping. (E): intraoperative Doppler ultrasound demonstrating marked reduction in perfusion of the upper Pole and mid-region following vascular clamping. (F): intra-operative image of the contralateral radical nephrectomy. (G): contrast-enhanced CT scan confirming the absence of the right kidney and presence of vascular staples at the left renal artery site. *: anterior segmental branch. **: posterior segmental branch

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  Fig. 2. Porcine CKD model characterization: (A): longitudinal body-weight changes, expressed relative to baseline weight, over 15 months. Top: individual values, group means with 95% CI. Bottom: summary mean and effect size with 95% CI. (B): glomerular filtration rate normalized to baseline, over 15 months, plotted as in A. (C): post-mortem left kidney/body weight ratio. Left: individual values, group means with 95% CI. Right: distribution of effect size with 95% CI. Bottom: summary mean and effect size with 95% CI. (D): Representative post-mortem images of left kidneys, showing ccompensatory hypertrophy and cortical scarring at the upper Pole. Scale bar = 1 cm. (E): post-mortem heart/body-weight ratio plotted as in C. Effect size interpretation (absolute value): no effect: (0), very small: (0.0,0.2), small: [0.2,0.5), medium: [0.5,0.8), large: [0.8,1.2), very large: [1.2,2.0), and huge: [2.0,∞)

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  Fig. 3. Adventitial vasa vasorum quantification in CKD pigs: (A): schematic of the porcine arterial tree, illustrating the ascending aorta (aAo), bicarotid artery trunk (BCaT), left and right common carotid arteries (LCCA, RCCA), and left and right iliac arteries (LIA, RIA). (B): representative contrast-enhanced ultrasound (CEUS) image of the LCCA, with adventitial vasa vasorum highlighted. (C): longitudinal assessment of carotid vasa vasorum, normalized to baseline over 15 months. Top: individual values, group means with 95% CI. Bottom: summary mean and effect size with 95% CI. (D): longitudinal assessment of iliac artery vasa vasorum, plotted as in C. Effect size interpretation (absolute value): no effect: (0), very small: (0.0,0.2), small: [0.2,0.5), medium: [0.5,0.8), large: [0.8,1.2), very large: [1.2,2.0), and huge: [2.0,∞)

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  Fig. 4. Visualization and quantification of vascular calcifications in CKD pigs: (A): contrast-free coronal CT reconstructions of the thoracoabdominal region. Vascular calcification foci are highlighted in the abdominal aorta (upper inset) and iliac arteries (lower inset). (B): axial CT slice of the abdominal aorta in CKD pigs, showing calcific deposits (circled). (C-F) quantification of arterial calcifications at 15 months in four vascular territories: common carotid artery (C), thoracic aorta (D), abdominal aorta (E), and iliac artery (F). For each panel, left plot displays individual values, group means with 95% CI. Right plots show distribution of effect size with 95% CI. Below each pair of plots, a summary of mean and effect size with 95% CI is provided. Effect size interpretation (absolute value): no effect: (0), very small: (0.0,0.2), small: [0.2,0.5), medium: [0.5,0.8), large: [0.8,1.2), very large: [1.2,2.0), and huge: [2.0,∞)

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  Fig. 5. Atheromatous plaque characterization in CKD pigs: (A): Representative adjacent sections of the abdominal aorta and coronary arteries stained with Masson’s trichome (left) and Von Kossa (right). Scale bar = 2.5 mm. (B): schematic of morphometric areas used for quantification. Four concentric regions were delineated on trichome-stained sections: area A: between the border of the adventitia and surrounding tissue; area M: between the border of the media layer and the adventitia; area I: between the media layer and the intima layer, following the lamina media; and area L: between the intima layer and the lumen. The specific areas were calculated indirectly: intima/plaque area = area I - Area L; media layer area = area M - Area I; adventitia area = area a – area M

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  Fig. 6. CKD-specific lipidomic signature: a total of 752 lipid species were quantified across serum and multiple tissues. Effect sizes with 99% CI confidence intervals were calculated to compare metabolite concentrations between control (CTRL) and CKD animals. Lipids exhibiting significant differences in serum and in at least one atheromatous lesion (fatty streak or atheroma plaque in aortic arch, iliac or carotid arteries), with consistent directionality, were selected. The abundance of these candidate metabolites was then explored in liver, adipose tissue, muscle, urine and kidney using the same criteria