Imaging of Dysfunctional Elastogenesis in Atherosclerosis Using an Improved Gadolinium-Based Tetrameric MRI Probe Targeted to Tropoelastin.

Federico Capuana,Begoña Lavin,Rachele Stefania,Sergio Padovan,Giuseppe Digilio,Eyad Almouazen,René M Botnar,Silvio Aime,Sara Lacerda,Yves Chevalier,Alkystis Phinikaridou,Laurence Heinrich-Balard

doi:10.1021/acs.jmedchem.1c01286

Abstract

Dysfunctional elastin turnover plays a major role in the progression of atherosclerotic plaques. Failure of tropoelastin cross-linking into mature elastin leads to the accumulation of tropoelastin within the growing plaque, increasing its instability. Here we present Gd4-TESMA, an MRI contrast agent specifically designed for molecular imaging of tropoelastin within plaques. Gd4-TESMA is a tetrameric probe composed of a tropoelastin-binding peptide (the VVGS-peptide) conjugated with four Gd(III)-DOTA-monoamide chelates. It shows a relaxivity per molecule of 34.0 ± 0.8 mM–1 s–1 (20 MHz, 298 K, pH 7.2), a good binding affinity to tropoelastin (KD = 41 ± 12 μM), and a serum half-life longer than 2 h. Gd4-TESMA accumulates specifically in atherosclerotic plaques in the ApoE–/– murine model of plaque progression, with 2 h persistence of contrast enhancement. As compared to the monomeric counterpart (Gd-TESMA), the tetrameric Gd4-TESMA probe shows a clear advantage regarding both sensitivity and imaging time window, allowing for a better characterization of atherosclerotic plaques.

Highlights

Atherosclerotic plaque rupture is one of the leading causes of life-threatening cardiovascular events
The general synthetic approach to Gd4-TESMA relied on the synthesis of a tetrameric Gd-DOTA-like chelate functionalized with a maleimide function
This gadolinium tetramer was conjugated to a cysteine residue added at the N-terminus of the tropoelastin specific binding sequence (C-VVGSPSAQDEASPLS) through the thiol/maleimide reaction to yield the final Gd4-TESMA imaging probe (Figure 1)

Summary

Introduction

Atherosclerotic plaque rupture is one of the leading causes of life-threatening cardiovascular events. The size of atherosclerotic plaques and the extent of luminal stenosis can be assessed by several imaging techniques, but these morphological parameters alone are not reliable diagnostic indicators of the risk for plaque rupture.[1] Depending on the morphology, composition, and histopathologic features, plaques have been classified as stable or high risk (vulnerable/unstable). Pathology studies revealed that plaques at high risk of rupturing with ensuing thrombosis are characterized by a thin fibrous cap, a large necrotic lipid-rich core, microcalcifications, and increased macrophage infiltration.[2] Increased plaque microvessel density may lead to intraplaque hemorrhage, further increasing the risk of rupture.[3] Phenotyping of atherosclerotic plaques by noninvasive imaging techniques has emerged as an essential requirement for risk assessment.[4] Vulnerable-plaque imaging aims at detecting biomarkers that would allow classifying plaques according to their risk of rupture

Results

Discussion

Conclusion