Scavenger Receptor Cysteine-Rich domains of Lysyl Oxidase-Like2 regulate endothelial ECM and angiogenesis through non-catalytic scaffolding mechanisms

Claudia Umana-Diaz (1, 2, 3) , Cathy Pichol-Thievend (1, 2, 3) , Marion F Marchand (1, 2, 3) , Yoann Atlas (1, 2, 3) , Romain Salza (4, 5) , Marilyne Malbouyres (6, 7) , Alain Barret (1, 2, 3) , Jérémie Teillon (8) , Corinne Ardidie-Robouant (1, 2, 3) , Florence Ruggiero (6, 7) , Catherine Monnot (1, 2, 3) , Philippe Girard (9) , Christophe Guilluy (10) , Sylvie Ricard-Blum (4, 5) , Stéphane Germain (2) , Laurent Muller (2)

1 PVM (UMR_7151) - Pathologie et virologie moléculaire
2 CIRB - Centre interdisciplinaire de recherche en biologie
3 INSERM U36 - Pathologie vasculaire et endocrinologie rénale - Chaire de médecine expérimentale
4 ICBMS - Institut de Chimie et Biochimie Moléculaires et Supramoléculaires
5 ASPE - Assemblages Supramoléculaires Péricellulaires et Extracellulaires
6 IGFL - Institut de Génomique Fonctionnelle de Lyon
7 Inflammasome NLRP3 – NLRP3 Inflammasome
8 CdF (institution) - Collège de France
9 IMM - Institut Mutualiste de Montsouris
10 IAB - Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble)

Lysyl oxidases are major actors of microenvironment and extracellular matrix (ECM) remodeling. These cross-linking enzymes are thus involved in many aspects of physiopathology, including tumor progression, fibrosis and cardiovascular diseases. We have already shown that Lysyl Oxidase-Like 2 (LOXL2) regulates collagen IV deposition by endothelial cells and angiogenesis. We here provide evidence that LOXL2 also affects deposition of other ECM components, including fibronectin, thus altering structural and mechanical properties of the matrix generated by endothelial cells. LOXL2 interacts intracellularly and directly with collagen IV and fibronectin before incorporation into ECM fibrillar structures upon exocytosis, as demonstrated by TIRF time-lapse microscopy. Furthermore, surface plasmon resonance experiments using recombinant scavenger receptor cysteine-rich (SRCR) domains truncated for the catalytic domain demonstrated their direct binding to collagen IV. We thus used directed mutagenesis to investigate the role of LOXL2 catalytic domain. Neither enzyme activity nor catalytic domain were necessary for collagen IV deposition and angiogenesis, whereas the SRCR domains were effective for these processes. Finally, surface coating with recombinant SRCR domains restored deposition of collagen IV by LOXL2-depleted cells. We thus propose that LOXL2 SRCR domains orchestrate scaffolding of the vascular basement membrane and angiogenesis through interactions with collagen IV and fibronectin, independently of the enzymatic crosslinking activity.

Domains: Life Sciences [q-bio]

Claudia Umana-Diaz, Cathy Pichol-Thievend, Marion F Marchand, Yoann Atlas, Romain Salza, et al.. Scavenger Receptor Cysteine-Rich domains of Lysyl Oxidase-Like2 regulate endothelial ECM and angiogenesis through non-catalytic scaffolding mechanisms. Matrix Biology, 2023, 88, pp.33 - 52. ⟨10.1016/j.matbio.2019.11.003⟩. ⟨hal-04029951⟩