Chade, Alejandro

Cardiovascular-kidney-metabolic (CKM) syndrome is a complex disease that affects up to 90% of US adults. This work characterized the renal transcriptomic landscape of a novel model of CKM in aging swine. The cardiovascular, renal, and metabolic parameters in pigs are very similar to those in humans, and our work may guide future studies aimed at modulating specific age-related renal abnormalities in CKM to slow disease progression and reduce mortality.

Scattered tubular-like cells (STCs) are dedifferentiated renal tubular cells that repair other damaged kidney cells. This study shows that renovascular disease (RVD) induces site-specific 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) modifications in DNA associated with transcriptional changes and impaired proliferative capacity of swine STCs, which could be mitigated in RVD-STCs treated with an epigenetic modulator. Therefore, our observations may contribute to the development of novel approaches to preserve the reparative capacity of STCs in individuals with RVD.

Background: Cardiovascular-Kidney-Metabolic (CKM) syndrome entails pathophysiological interactions leading to multiorgan dysfunction and a high rate of cardiovascular morbidity and mortality, largely driven by the development of heart failure (HF). Mechanisms leading to HF in CKM syndrome are unknown. We created a model of CKM syndrome in swine and showed that cardiac abnormalities associate with cardiac microvascular (MV) rarefaction and decreased expression and availability of VEGF. Emerging research focuses on micro-RNAs (miRNAs) and their role in the pathophysiology of HF, but whether they play a role in the development of cardiac injury in CKM syndrome via VEGF modulation has not been investigated. We aimed to identify specific miRNAs that can target VEGF to test the hypothesis that their direct cardiac inhibition will rescue cardiac VEGF, improve angiogenic signaling, and cardiac hemodynamics in CKM syndrome. Methods: CKM (bilateral renal artery stenosis+high-cholesterol/high-carbohydrate feeding) and normal pigs (n=4 each, 2 males/2 females) were studied for 14 weeks. Unbiased micro-RNA sequencing identified 26 miRNAs differentially expressed in CKM pig hearts, of which only miR-452-5p can target VEGFA, which was confirmed by pulldown analysis and qPCR. In addition, CKM (n=4, 2 males/2 females) were treated after 6 weeks with a single intracoronary (IC) administration of a miR-452-5p inhibitor (antagomiR). Cardiac hemodynamics (echocardiography), MV architecture (3D micro-CT) and cardiac expression of VEGF and miR-452-5p (RT-qPCR) were quantified before IC miR-452-5p inhibition, and again at the end of the study (14 weeks). Results: Pigs with CKM syndrome showed blunted diastolic relaxation, increased cardiac remodeling, and reduced subendocardial MV density and cardiac VEGF expression, accompanied by higher cardiac expression of miR-452-5p compared to normal pigs. Notably, most of these parameters were improved after in vivo IC miR-452-5p inhibition (single dose). Blood pressure was unchanged ( Table 1 ). Conclusions: Our study supports a role for VEGF in preserving cardiac MV integrity in CKM syndrome. Also, it identifies a targeted post-transcriptional regulation of VEGF via miR-452-5p and supports the feasibility and efficacy of its mechanistic modulation in vivo. The predictive power of our model conceptually supports the potential of a novel targeted intervention to protect the heart in CKM syndrome. [Table: see text] Support: NIH AG084154 (ARC), and DK129240 (AE) This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

We provide the first longitudinal and most comprehensive analysis of the intrarenal microvasculature in a slowly progressive model of autosomal dominant polycystic kidney disease (ADPKD) and integrate the findings with studies in a young cohort of individuals with ADPKD. We identified vasculature-related pathways that could be targeted for therapeutic interventions and contribute promising, noninvasive biomarkers in patients with ADPKD. Alterations of the intrarenal microcirculation may affect drug delivery; a better understanding of its longitudinal changes may aid in treatment management.

Abstract Background and Aims Almost 36 million US adults (1 in 7, 14%) suffer from chronic kidney disease (CKD), and precision strategies to offset the progression to end-stage kidney disease are in dire need. We took advantage of a novel preclinical translational model of CKD in swine and single nucleus RNA sequencing (snRNA-seq) to identify potential therapeutic targets via characterization of the renal cell-specific transcriptomic landscape. Method Normal and CKD pigs were euthanized after 14 weeks (n = 3/group), kidneys were harvested, nuclei isolated, libraries prepared, and snRNA-seq performed. UMAP plots were generated with Seurat (v4.4.0) using default parameters. The expression profile of marker genes across single nuclei clusters was visualized to define renal cell types and ranked based on the number of differentially expressed genes (DEGs). The top cell type was subjected to subsequent analysis and DEG sorted based on their involvement in angiogenesis, inflammation, and fibrosis, which are prominent injury pathways in the pathophysiology of CKD. To mechanistically study the role of top overlapping candidate DEGs and support their potential as therapeutic targets, in vitro studies in primary renal vascular endothelial cells (RECs) were performed. Results A total of 39,935 nuclei were analyzed. Thirty clusters were identified and filtered by canonical gene markers, revealing 18 unique renal cell types (Fig. 1A). Endothelial cells exhibited the highest number of DEGs (Fig. 1B). Volcano plots (Fig. 1C) and Venn diagram (Fig. 1D) analysis of endothelial cells identified unique DEGs among three main renal injury pathways in CKD (angiogenesis, inflammation, fibrosis), with 2 overlapping DEGs (VWF, LAMA3) upregulated and participating in inflammation and fibrosis. Targeted silencing of VWF and LAMA3 (randomly selected) in RECs using siRNA ameliorated C5b9 and TGF-ẞ immunoreactivity, suggesting attenuated inflammatory and fibrogenic activity, respectively (Fig. 1E). Conclusion Our work characterized the single-nuclear renal cell transcriptomic landscape of a translational model of CKD and identified endothelial cells as the cluster with the largest number of DEGs. Our observations support the candidacy of VWF and LAMA3 as potential targets to decrease renal endothelial cell inflammation and fibrosis, which could pave the way for new therapeutic strategies in patients with CKD.

Abstract Background and Aims Almost 36 million US adults (1 in 7, 14%) suffer from chronic kidney disease (CKD), which is more prevalent in the elderly, affecting up to 34% of adults ≥ over 65. CKD has limited therapeutic options, partly due to the lack of large animal models that allow the translational study of the impact of age on CKD. This work reports the first preclinical model of CKD in naturally aged swine. Method CKD was induced in naturally aged (6-9 years old) and young (3 months old) domestic pigs (bilateral renal artery stenosis + high-cholesterol/high-carbohydrate diet), followed for 14 weeks, and compared to normal controls (n = 4–5/group). Renal hemodynamics were quantified in vivo by multidetector CT. Metabolic parameters, renal microvascular (MV) architecture (3D-micro-CT), and morphometric analysis (staining) were investigated ex vivo. Results Both young and old pigs developed CKD, but the renal and metabolic phenotype was accentuated in aging animals. We found that superimposition of aging on CKD resulted in higher triglycerides, glucose levels, insulin resistance, and adipose tissue accumulation (subcutaneous, visceral, perirenal) (Fig. 1A and B), accompanied by a more severe renal dysfunction (C), cortical MV rarefaction, and reduced peritubular capillary density (D and E), and more severe structural damage disclosed by increased glomerulosclerosis, tubule-interstitial fibrosis, and tubular injury (F). Notably, the severity of renal MV rarefaction in aging CKD correlated with functional and structural abnormalities (simple linear regression analysis, G). Conclusion Our study establishes the first large animal model of aging CKD, allowing the investigation of age as a biological variable in kidney-metabolic diseases. Age exacerbates metabolic and renal abnormalities, and our data support the critical role of renal MV disease in aging CKD and suggest that its exacerbation is a potential key player in its progression. This novel translational platform could foster new studies toward the elucidation of novel mechanisms of age-related deterioration of renal function in CKD and for developing therapeutic interventions.

Abstract Background and Aims Patients with chronic kidney disease (CKD) have a higher risk of developing heart failure with preserved ejection fraction (HFpEF). Growing evidence supports that changes in cardiac microcirculation are a major pathophysiological driver of HFpEF. We created a model of CKD stage 3 that subsequently developed a cardiac phenotype with numerous traits of HFpEF (CKD-HFpEF), including microvascular (MV) rarefaction and decreased availability of the pro-angiogenic growth factor vascular endothelial growth factor (VEGF) in the heart. We hypothesize that cardiac VEGF therapy will attenuate MV injury and improve cardiac function in CKD-HFpEF. Method CKD and normal pigs (n = 6 each, 3 males/3 females) were studied for 14 weeks. In addition, CKD-HFpEF pigs (n = 6, 3 males/3 females) were treated after 6 weeks of disease with a single intracoronary (IC) administration of VEGF attached to a drug-delivery vector (ELP-VEGF) that increases cardiac targeting, and effects quantified 8 weeks later. Renal (multi-detector CT) and cardiac (echocardiography, PV loops) morphology and function were quantified in vivo, microvascular (MV) density (3D micro-CT) ex vivo, and cardiac expression of VEGF was quantified by qPCR and western blot, as VEGF-related genes by bulk mRNA-sequencing (n = 3/group). Results A single IC administration of ELP-VEGF improved MV density and rescued cardiac function compared to placebo similarly in male and female pigs and independent of hypertension (Fig. 1A–C). These improvements were accompanied by improved mRNA and protein expression of VEGF and restored expression of VEGF-related pro- and anti-angiogenic genes (Fig. 1D and E). Conclusion Our study supports the prominent role of cardiac MV disease in the development of HFpEF in CKD, supports the critical need for healthy VEGF signaling to preserve the heart in CKD, and shows the feasibility of a potential new therapeutic strategy via a targeted mechanistic intervention.

Chronic kidney disease (CKD), heart failure with preserved ejection fraction (HFpEF), and the recently defined cardiovascular-kidney-metabolic (CKM) syndrome are important challenges for the health care system, associated with a staggering increase in morbidity and mortality rates. Unfortunately, the complex pathophysiology of these processes remains to be fully elucidated, imposing hurdles to prevent their development and decelerate their progression. Large animal models are critical for understanding cardio-renal pathophysiology and for the discovery of new therapies. Among them, pig models offer significant translational power to grasp cardio-renal pathophysiology and to test the feasibility and efficacy of new strategies. We recently developed and characterized swine models of CKD, HFpEF, and CKM syndrome that closely recapitulate human disease and allow for the study of the impact of biological variables, such as sex or age, on these conditions. Furthermore, data from these studies revealed key mechanisms implicated in the pathogenesis of CKD, HFpEF, and CKM syndrome and identified potential targets for intervention. This focused mini review highlights the main features of these novel swine models and discusses ongoing and future research aimed at developing novel therapies.