DDR1 Regulates Femoral Arterial Calcification in Lower‐Extremity Artery Disease Through NF ‐Kappa B Activation

Acta Physiologica, vol. 242, pp. e70146

Abstract

AIM: Lower-extremity arterial disease (LEAD) is a manifestation of atherosclerotic cardiovascular disease, affecting 230 million people worldwide with increasing prevalence. Medial arterial calcification (MAC) is common in LEAD patients and contributes to disease-related mortality. However, therapeutic strategies targeting femoral MAC are lacking, and its underlying mechanisms remain unclear. This study aimed to identify molecular drivers of femoral MAC in LEAD. METHODS & RESULTS: Calcium deposits and pro-calcifying markers were analyzed in human patient samples using von Kossa staining, immunofluorescence, and gene expression analysis. Femorals showed significantly more calcification and pro-calcifying gene expression than carotids. Given MAC abundance in LEAD, we assessed medial calcification in Apoe-/- mice fed a WD for 4/21 weeks. Digital PCR revealed upregulation of Ddr1 and Bmp2 in femoral versus carotid arteries after 21 weeks of WD. DDR1 expression positively correlated with calcification in human femoral samples. In vitro experiments with mouse femoral vs. carotid vascular smooth muscle cells (VSMCs) confirmed a significantly higher prevalence of calcifying proteins (DDR1, BMP2, and RUNX2) in femoral VSMCs. Additionally, calcification analyses in murine and human VSMCs showed that DDR1 inhibition reduced, while DDR1 activation increased, calcium deposition. Transcriptomic analysis revealed elevated NF-κB expression in human femoral arteries, matching data in femoral VSMCs. DDR1 stimulation activated NF-κB, and its inhibition blocked DDR1-induced calcification. CONCLUSION: This study identifies DDR1 as a key driver of calcification in LEAD, operating through NF-κB activation and the expression of calcifying proteins. Targeting DDR1 may offer a novel therapeutic approach to prevent MAC in LEAD.

Authors 14

1. Manovriti Thakur

   University of Bern · University Hospital of Bern

   Affiliation as printed

   Department for BioMedical Research (DBMR) University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

2. Thibaut Quillard

   Centre National de la Recherche Scientifique · Inserm · Centre Hospitalier Universitaire de Nantes · Institut du Thorax · Nantes Université

   Affiliation as printed

   Nantes Université, CHU Nantes, CNRS, INSERM, L'institut du Thorax Nantes France

3. Nico Angliker

   University of Bern · University Hospital of Bern

   Affiliation as printed

   Department for BioMedical Research (DBMR) University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

4. Mark Siegrist

   University of Bern · University Hospital of Bern

   Affiliation as printed

   Department for BioMedical Research (DBMR) University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital University of Bern Bern Switzerland

   Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

5. Yvonne J. F. M. Jansen

   Ludwig-Maximilians-Universität München · Institut für Prophylaxe und Epidemiologie der Kreislaufkrankheiten

   Affiliation as printed

   Institute for Cardiovascular Prevention (IPEK) Ludwig‐Maximilians‐University Munich (LMU) Munich Germany

   Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich (LMU), Munich, Germany

6. Yi Yan

   Shanghai Jiao Tong University · Shanghai Children's Medical Center

   Affiliation as printed

   Heart Center and Shanghai Institute of Pediatric Congenital Heart Disease, Shanghai Children's Medical Center, National Children's Medical Center Shanghai Jiao Tong University School of Medicine Shanghai China

   Heart Center and Shanghai Institute of Pediatric Congenital Heart Disease, Shanghai Children's Medical Center, National Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China

7. Julia Wollenhaupt Aachen Institute for Molecular Cardiovascular Research

   RWTH Aachen University · Universitätsklinikum Aachen · Maastricht University

   Affiliation as printed

   Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM) Maastricht University Maastricht the Netherlands

   Institute for Molecular Cardiovascular Research (IMCAR) Uniklinik Aachen, RWTH Aachen University Aachen Germany

   Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

   Institute for Molecular Cardiovascular Research (IMCAR), Uniklinik Aachen, RWTH Aachen University, Aachen, Germany

8. Claudia Goettsch Aachen Department of Internal Medicine I (Cardiology)

   RWTH Aachen University · Universitätsklinikum Aachen · Technische Universität Dresden · University Hospital Carl Gustav Carus

   Affiliation as printed

   Department of Internal Medicine I University Hospital of the RWTH Aachen Aachen Germany

   Institute of Physiology, Faculty of Medicine Carl Gustav Carus Technical University Dresden Dresden Germany

   Department of Internal Medicine I, University Hospital of the RWTH Aachen, Aachen, Germany

   Institute of Physiology, Faculty of Medicine Carl Gustav Carus, Technical University Dresden, Dresden, Germany

9. Lars Mäegdefessel

   German Centre for Cardiovascular Research · Technical University of Munich · TUM Klinikum

   Affiliation as printed

   DZHK (German Centre for Cardiovascular Research) Partner Site Munich Heart Alliance Munich Germany

   Department for Vascular and Endovascular Surgery, TUM Klinikum Technical University Munich Munich Germany

   DZHK (German Centre for Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany

   Department for Vascular and Endovascular Surgery, TUM Klinikum, Technical University Munich, Munich, Germany

10. Nadja Sachs

    German Centre for Cardiovascular Research · Technical University of Munich · TUM Klinikum

    Affiliation as printed

    DZHK (German Centre for Cardiovascular Research) Partner Site Munich Heart Alliance Munich Germany

    Department for Vascular and Endovascular Surgery, TUM Klinikum Technical University Munich Munich Germany

    DZHK (German Centre for Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany

    Department for Vascular and Endovascular Surgery, TUM Klinikum, Technical University Munich, Munich, Germany

11. Marc Schindewolf

    University of Bern · University Hospital of Bern

    Affiliation as printed

    Department for BioMedical Research (DBMR) University of Bern Bern Switzerland

    Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital University of Bern Bern Switzerland

    Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

12. Drosos Kotelis

    University Hospital of Bern

    Affiliation as printed

    Department of Vascular Surgery University Hospital Bern Bern Switzerland

    Department of Vascular Surgery, University Hospital Bern, Bern, Switzerland

13. Heidi Noels Aachen Institute for Molecular Cardiovascular Research

    RWTH Aachen University · Universitätsklinikum Aachen · Maastricht University

    Affiliation as printed

    Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM) Maastricht University Maastricht the Netherlands

    Institute for Molecular Cardiovascular Research (IMCAR) Uniklinik Aachen, RWTH Aachen University Aachen Germany

    Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

    Institute for Molecular Cardiovascular Research (IMCAR), Uniklinik Aachen, RWTH Aachen University, Aachen, Germany

14. Yvonne Döring corresponding

    Ludwig-Maximilians-Universität München · German Centre for Cardiovascular Research · University of Bern · University Hospital of Bern · Institut für Prophylaxe und Epidemiologie der Kreislaufkrankheiten

    Affiliation as printed

    DZHK (German Centre for Cardiovascular Research) Partner Site Munich Heart Alliance Munich Germany

    Department for BioMedical Research (DBMR) University of Bern Bern Switzerland

    Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital University of Bern Bern Switzerland

    Institute for Cardiovascular Prevention (IPEK) Ludwig‐Maximilians‐University Munich (LMU) Munich Germany

    DZHK (German Centre for Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany

    Division of Angiology, Swiss Cardiovascular Center, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland

    Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich (LMU), Munich, Germany