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1-Phosphatidylinositol 3-Phosphate 5-Kinase Inhibition by Apilimod Promotes an Adipocyte-Like Vascular Smooth Muscle Cell Phenotype and Prevents Arterial Calcification

Hense, Nicolas ; Gorgels, Andrea ; Mir, Bilal ; van der Vorst, Emiel P.C. ; Falgayrac, Guillaume ; Terliesner, Mara ; Goncalves, Isabel LU orcid ; Matic, Ljubica ; Edsfeldt, Andreas LU orcid and Sun, Jiangming LU orcid , et al. (2026) In Circulation Research 138(3).
Abstract

BACKGROUND: Cardiovascular calcification is a significant predictor and contributor to cardiovascular diseases. Vascular smooth muscle cell (SMC)-derived extracellular vesicles (EVs) play a crucial role in microcalcification formation. EVs can originate from the endosomal system, and PIKFYVE (1-phosphatidylinositol 3-phosphate 5-kinase), a lipid kinase, plays a key role in endomembrane maturation. We hypothesize that PIKFYVE inhibition will modulate EV cargo and, thereby, arterial calcification. METHODS: Human coronary artery SMCs were cultured in osteogenic media to induce calcification. PIKFYVE inhibition was achieved using the pharmacological inhibitor apilimod and siRNA. We characterized the SMC phenotype and EVs through proteomics,... (More)

BACKGROUND: Cardiovascular calcification is a significant predictor and contributor to cardiovascular diseases. Vascular smooth muscle cell (SMC)-derived extracellular vesicles (EVs) play a crucial role in microcalcification formation. EVs can originate from the endosomal system, and PIKFYVE (1-phosphatidylinositol 3-phosphate 5-kinase), a lipid kinase, plays a key role in endomembrane maturation. We hypothesize that PIKFYVE inhibition will modulate EV cargo and, thereby, arterial calcification. METHODS: Human coronary artery SMCs were cultured in osteogenic media to induce calcification. PIKFYVE inhibition was achieved using the pharmacological inhibitor apilimod and siRNA. We characterized the SMC phenotype and EVs through proteomics, transcriptomics, and kinomics. Ldlr-deficient mice fed a high-fat, high-cholesterol diet received apilimod for 5 weeks. RESULTS: Calcified human arteries and SMCs exhibited increased PIKFYVE protein expression compared with controls. In calcifying SMCs, phosphatidylinositol 3-phosphate levels were reduced but restored by apilimod. Apilimod prevented matrix mineralization and collagen deposition in calcifying SMCs, accompanied by reduced procollagen 1A1 secretion. Apilimod inhibited TNAP (tissue-nonspecific alkaline phosphatase) at mRNA, protein, and activity levels. EVs released from apilimod-treated calcifying SMCs exhibited lower mineral cargo, reduced aggregation potential, and diminished TNAP cargo. Phenotypic omics analyses revealed that apilimod induced a shift toward an adipocyte-like SMC phenotype, marked by upregulation of adipogenic TFs (transcription factors), fatty acid metabolism genes, and increased fatty acid uptake. Reactivation of YAP (Yes-associated protein)-TEAD (transcriptional enhancer factor) signaling partially reversed these phenotypic changes. In vivo, apilimod reduced vascular calcification and plaque TNAP activity but increased plaque lipid accumulation. CONCLUSIONS: Pharmacological inhibition of PIKFYVE disrupts YAP-TEAD signaling, thereby reducing arterial calcification by limiting the calcification potential of EVs and suppressing osteogenic SMC programming, but also induces an adipocyte-like SMC phenotype with lipid accumulation. These findings emphasize the need to consider SMC phenotypic plasticity and potential adverse effects when developing therapeutic strategies for arterial calcification.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
amorphous calcium phosphate, chromatography, liquid, extracellular vesicles, muscle, smooth, vascular, phenotype
in
Circulation Research
volume
138
issue
3
article number
e326772
publisher
Lippincott Williams & Wilkins
external identifiers
  • pmid:41533526
  • scopus:105029043689
ISSN
0009-7330
DOI
10.1161/CIRCRESAHA.125.326772
language
English
LU publication?
yes
id
1d7a5992-f7a9-487d-ab6e-30ae87816fb2
date added to LUP
2026-02-19 13:11:35
date last changed
2026-02-20 03:20:16
@article{1d7a5992-f7a9-487d-ab6e-30ae87816fb2,
  abstract     = {{<p>BACKGROUND: Cardiovascular calcification is a significant predictor and contributor to cardiovascular diseases. Vascular smooth muscle cell (SMC)-derived extracellular vesicles (EVs) play a crucial role in microcalcification formation. EVs can originate from the endosomal system, and PIKFYVE (1-phosphatidylinositol 3-phosphate 5-kinase), a lipid kinase, plays a key role in endomembrane maturation. We hypothesize that PIKFYVE inhibition will modulate EV cargo and, thereby, arterial calcification. METHODS: Human coronary artery SMCs were cultured in osteogenic media to induce calcification. PIKFYVE inhibition was achieved using the pharmacological inhibitor apilimod and siRNA. We characterized the SMC phenotype and EVs through proteomics, transcriptomics, and kinomics. Ldlr-deficient mice fed a high-fat, high-cholesterol diet received apilimod for 5 weeks. RESULTS: Calcified human arteries and SMCs exhibited increased PIKFYVE protein expression compared with controls. In calcifying SMCs, phosphatidylinositol 3-phosphate levels were reduced but restored by apilimod. Apilimod prevented matrix mineralization and collagen deposition in calcifying SMCs, accompanied by reduced procollagen 1A1 secretion. Apilimod inhibited TNAP (tissue-nonspecific alkaline phosphatase) at mRNA, protein, and activity levels. EVs released from apilimod-treated calcifying SMCs exhibited lower mineral cargo, reduced aggregation potential, and diminished TNAP cargo. Phenotypic omics analyses revealed that apilimod induced a shift toward an adipocyte-like SMC phenotype, marked by upregulation of adipogenic TFs (transcription factors), fatty acid metabolism genes, and increased fatty acid uptake. Reactivation of YAP (Yes-associated protein)-TEAD (transcriptional enhancer factor) signaling partially reversed these phenotypic changes. In vivo, apilimod reduced vascular calcification and plaque TNAP activity but increased plaque lipid accumulation. CONCLUSIONS: Pharmacological inhibition of PIKFYVE disrupts YAP-TEAD signaling, thereby reducing arterial calcification by limiting the calcification potential of EVs and suppressing osteogenic SMC programming, but also induces an adipocyte-like SMC phenotype with lipid accumulation. These findings emphasize the need to consider SMC phenotypic plasticity and potential adverse effects when developing therapeutic strategies for arterial calcification.</p>}},
  author       = {{Hense, Nicolas and Gorgels, Andrea and Mir, Bilal and van der Vorst, Emiel P.C. and Falgayrac, Guillaume and Terliesner, Mara and Goncalves, Isabel and Matic, Ljubica and Edsfeldt, Andreas and Sun, Jiangming and Hedin, Ulf and Hayat, Sikander and Thiel, Jessica and Ramirez-Torres, Maria Alejandra and Barsoum, Mirna and Rauch, Alexander and Rauner, Martina and Gombert, Alexander and Preisinger, Christian and Noels, Heidi and Marx, Nikolaus and Goettsch, Claudia}},
  issn         = {{0009-7330}},
  keywords     = {{amorphous calcium phosphate; chromatography, liquid; extracellular vesicles; muscle, smooth, vascular; phenotype}},
  language     = {{eng}},
  number       = {{3}},
  publisher    = {{Lippincott Williams & Wilkins}},
  series       = {{Circulation Research}},
  title        = {{1-Phosphatidylinositol 3-Phosphate 5-Kinase Inhibition by Apilimod Promotes an Adipocyte-Like Vascular Smooth Muscle Cell Phenotype and Prevents Arterial Calcification}},
  url          = {{http://dx.doi.org/10.1161/CIRCRESAHA.125.326772}},
  doi          = {{10.1161/CIRCRESAHA.125.326772}},
  volume       = {{138}},
  year         = {{2026}},
}