Matrix vesicle biomimetics harboring Annexin A5 and alkaline phosphatase bind to the native collagen matrix produced by mineralizing vascular smooth muscle cells

Author(s)

Bolean, Mayte ^[1] ; Izzi, Benedetta ^[2] ; van Kerckhoven, Soetkin ^[3] ; Bottini, Massimo ^{[4, 5]} ; Ramos, Ana Paula ^[1] ; Millan, Jose Luis ^[5] ; Hoylaerts, Marc F. ^[6] ; Ciancaglini, Pietro ^[1]

Affiliation

^[1] Univ Sao Paulo, FFCLRP USP, Dept Chem, Ribeirao Preto, SP - Brazil

^[2] IRCCS NEUROMED, Dept Epidemiol & Prevent, Pozzilli, IS - Italy

^[3] UZ Leuven Gasthuisberg, Functiemetingen Pneumol, Leuven - Belgium

^[4] Univ Roma Tor Vergata, Dept Expt Med & Surg, Rome - Italy

^[5] Sanford Burnham Prebys Med Discovery Inst, La Jolla, CA - USA

^[6] Univ Leuven, Ctr Mol & Vasc Biol, Dept Cardiovasc Sci, Leuven - Belgium

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1864, n. 8 AUG 2020.
• Web of Science Citations: 0

Abstract

Backgound: Vascular smooth muscle cells (VSMCs) transdifferentiated ectopically trigger vascular calcifications, contributing to clinical cardiovascular disease in the aging population. AnxA5 and TNAP play a crucial role in (patho)physiological mineralization. Methods: We performed affinity studies between DPPC and 9:1 DPPC:DPPS-proteoliposomes carrying AnxA5 and/or TNAP and different types of collagen matrix: type I, II, I + III and native collagenous extracellular matrix (ECM) produced from VSMCs with or without differentiation, to simulate ectopic calcification conditions. Results: AnxA5-proteoliposomes had the highest affinity for collagens, specially for type II. TNAP-proteoliposomes bound poorly and the simultaneous presence of TNAP in the AnxA5-proteoliposomes disturbed interactions between AnxA5 and collagen. DPPC AnxA5-proteoliposomes affinities for ECM from transdifferentiating cells went up 2-fold compared to that from native VSMCs. The affinities of DPPC:DPPS-proteoliposomes were high for ECM from VSMCs with or without differentiation, underscoring a synergistic effect between AnxA5 and DPPS. Co-localization studies uncovered binding of proteoliposomes harboring AnxA5 or TNAP+AnxA5 to various regions of the ECM, not limited to type II collagen. Conclusion: AnxA5-proteoliposomes showed the highest affinities for type II collagen, deposited during chondrocyte mineralization in joint cartilage. TNAP in the lipid/protein microenvironment disturbs interactions between AnxA5 and collagen. These findings support the hypothesis that TNAP is cleaved from the MVs membrane just before ECM binding, such facilitating MV anchoring to ECM via AnxA5 interaction. General significance: Proteoliposomes as MV biomimetics are useful in the understanding of mechanisms that regulate the mineralization process and may be essential for the development of novel therapeutic strategies to prevent or inhibit ectopic mineralization. (AU)

• FAPESP's process: 17/08892-9 - Bioactive surfaces designed from Langmuir-Blodgett Films and Biominerals
• Grantee: Ana Paula Ramos
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/00371-1 - Are the interactions between collagen and proteins/enzymes present in the matriz vesicles responsible for the control in the biomineralization process?
• Grantee: Maytê Bolean Correia
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 15/06814-5 - Do interactions between collagen and proteins/enzymes present in matrix vesicles control biomineralization?
• Grantee: Maytê Bolean Correia
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor