A Murine Model of Cardiovascular-Kidney-Metabolic Syndrome Demonstrates Compromised Limb Function in the Ischemic Hind Limb

Authors

• Saran Lotfollahzadeh
• Herreet Paul
• Joshua Bonifacio
• Ricardo Almiron
• Isaac Hockestra
• Kylla Przekop
• Trent Yamamoto
• Maria Carmen Piqueras
• Wenqing Yin
• Kashvi Sethuraman
• Asha Jose
• Marina Malikova
• Jeffrey J Siracuse
• Mostafa Belghasem
• Howard Cabral
• Nazish Sayed
• Vipul C Chitalia

Language

English

Publication Date

2-1-2026

Journal

Kidney360 .

DOI

10.34067/KID.0000000900

PMID

41082313

PMCID

PMC12935364

PubMedCentral® Posted Date

10-13-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Key Points:

1. Cardiovascular-kidney-metabolic (CKM) is a global public health problem; however, its mediators remain poorly known, in part due to the lack of a reliable animal model.

2. A combination of high fat and adenine diet recapitulate some of the CKD and metabolic phenotypes of CKM.

3. This model also demonstrates myocardial fibrosis and peripheral artery disease with sex-based differences and can be leveraged to probe mechanisms of CKM.

Background: Cardiovascular-kidney-metabolic (CKM) syndrome is a public health problem in the United States and results in premature cardiovascular disease at a relatively preserved GFR. The molecular mediators of CKM are poorly understood, partly due to the lack of a reliable animal model. We set out to generate an animal model with renal and metabolic dysfunctions, using peripheral artery disease (PAD) as a CKM manifestation.

Methods: C57BL/6 male and female mice were randomized into four groups: a normal diet (controls), a 0.2% adenine diet (AD, a CKD model), a high-fat diet (HFD, a metabolic model), and a combination of HFD+AD (a potential CKM model). The mice underwent a hind limb ischemia, followed by an array of structural, endurance, and postexercise hyperemia assays.

Results: Compared with control mice, HFD+AD male mice had 23%–50% higher weight and GFR than the AD group (P = 0.003). The kidneys of HFD+AD showed tubular atrophy, tubulointerstitial fibrosis, immune infiltration, glomerulomegaly, consistent with glomerular hyperperfusion, hypercholesterolemia, impaired glucose tolerance, and adipophilin in the liver, an early marker of hepatic steatosis, and myocardial fibrosis. The HFD+AD mice showed reductions in the hind limb perfusion ratios, microcapillary density, type 2 muscle fibers, and increased muscle fibrosis, immune infiltration, and lowest cross-sectional muscle area. Female CKM mice revealed distinct differences from male mice. Compared with AD and HFD alone, female CKM mice exposed to HFD+AD demonstrated additive phenotypes in endurance assays (distance traveled, exhaustion time, and grip strength) without a similar effect in postischemia perfusion, suggesting skeletal muscle, and microcapillary dysfunction.

Conclusions: A combination of HFD+AD in mice displays features of CKD, metabolic disorders, and cardiovascular disease at a higher GFR, consistent with human CKM. This model can be explored to probe the mechanisms and heterogeneity and sex-specific differences in CKM.

Keywords

cardiovascular disease, CKD, kidney disease

Published Open-Access

yes

Recommended Citation

Lotfollahzadeh, Saran; Paul, Herreet; Bonifacio, Joshua; et al., "A Murine Model of Cardiovascular-Kidney-Metabolic Syndrome Demonstrates Compromised Limb Function in the Ischemic Hind Limb" (2026). Faculty and Staff Publications. 3.
https://digitalcommons.library.tmc.edu/huffington_pub/3