Integrated single-cell RNA-seq analysis reveals mitochondrial calcium signaling as a modulator of endothelial-to-mesenchymal transition
Mathilde Lebas, Giorgia Chinigò, Evan Courmont, Louay Bettaieb, Amani Machmouchi, Jermaine Goveia, Aleksandar Beatovic, Job Van Kerckhove, Cyril Robil, Fabiola Silva Angulo, Mauro Vedelago, Alina Errerd, Lucas Treps, Vance Gao, Hilda Carolina Delgado-De la Herrán, Alicia Mayeuf-Louchart, Laurent L’homme, Mohamed Chamlali, Camille Dejos, Valérie Gouyer, Venkata Naga Srikanth Garikipati, Dhanendra Tomar, Hao Yin, Hajime Fukui, Stefan Vinckier, Anneke Stolte, Lena‐Christin Conradi, Fabrice Infanti, Loïc Lemonnier, Elisabeth M. Zeisberg, Yonglun Luo, Lin Lin, Jean‐Luc Desseyn, J. Geoffrey Pickering, Raj Kishore, Muniswamy Madesh, David Dombrowicz, Fabiana Perocchi, Bart Staels, Alessandra Fiorio, Dimitra Gkika, Anna Rita Cantelmo
Abstract
Endothelial cells (ECs) are highly plastic, capable of differentiating into various cell types. Endothelial-to-mesenchymal transition (EndMT) is crucial during embryonic development and contributes substantially to vascular dysfunction in many cardiovascular diseases (CVDs). While targeting EndMT holds therapeutic promise, understanding its mechanisms and modulating its pathways remain challenging. Using single-cell RNA sequencing on three in vitro EndMT models, we identified conserved gene signatures. We validated original regulators in vitro and in vivo during embryonic heart development and peripheral artery disease. EndMT induction led to global expression changes in all EC subtypes rather than in mesenchymal clusters. We identified mitochondrial calcium uptake as a key driver of EndMT; inhibiting mitochondrial calcium uniporter (MCU) prevented EndMT in vitro, and conditional Mcu deletion in ECs blocked mesenchymal activation in a hind limb ischemia model. Tissues from patients with critical limb ischemia with EndMT features exhibited significantly elevated endothelial MCU. These findings highlight MCU as a regulator of EndMT and a potential therapeutic target.