P1215Non-invasive angiogenesis assessment with PET/CT in a rabbit model of myocardial infarction: feasibility and comparison with molecular markers
Authors: Grandjean C E , Korshin A , Smerup M , Jensen J K , Follin B , Brandt-Larsen M , Pedersen S F , Ripa R S , Binderup T , Kjaer A 
Published: October 2019

Abstract

Background
Translational molecular imaging using Positron Emission Tomography/Computed Tomography (PET/CT) and Transesophageal Echocardiography (TEE) are powerful non-invasive tools for investigation and monitoring of cardiovascular functions. Following myocardial infarction (MI), formation of new blood vessels (angiogenesis) is of importance in the healing process and can be assessed with the PET tracer 68Ga-NODAGA-E[(cRGDyK)]2 (68Ga-RGD).

Purpose
The purpose of the study was to assess angiogenesis in a rabbit model of myocardial infarction using molecular imaging (PET/CT) and furthermore to validate the findings using TEE, immunohistochemistry (IHC) and quantitative PCR (qPCR).

Methods
Ten NZW rabbits were used (MI: n=5, SHAM: n=5). Angiogenesis was assessed with 68Ga-RGD before, 1 and 3 weeks after interventions using PET/CT (Inveon System, Siemens Health Care). TEE was used per-operatively and at termination for assessment of the MI (S8–3T probe, iE33 System, Phillips). Following the final scans, hearts were harvested for ex vivo analyses. Short axis slides were stained for collagen deposition and myocardial differentiation (H&E and Masson's trichrome), endothelial cells (CD31), and macrophage infiltration (RAM11). Gene expression alterations related to wound healing response (inflammation, granulation, and tissue remodeling) were measured using qPCR arrays (84 genes analyzed).

Results
One week after the interventions, 68Ga-RGD uptake, as assessed with PET was increased in the infarct area when compared to the remote zone of the same rabbit as well as compared to the SHAM group. Three weeks after intervention, there was no difference in 68Ga-RGD uptake between groups. High quality TEE images were obtained in all rabbits. Ex vivo analyses at 3 weeks after intervention revealed moderate vessel formation (CD31) in the infarct zone, none in the border zone, whereas surrounding viable myocardium had visible CD31 positive vessels comparable to the SHAM group. Macrophage infiltration (RAM11) and collagenous scaring (Masson's trichrome) was pronounced in the infarcted area. Gene expression alterations in the infarct area (30 of 84 genes upregulated) were dominated by increased expression of collagens (COL1A2, COL5A1, COL5A4), inflammatory chemokines and cytokines (CCL2, IL1A, IL1B, IL6, IL10) and ECM remodeling enzymes (uPAR, TPA, TIMP1, SERPINE1, MMP9) in the MI group compared to the SHAM group, whereas integrins involved in the angiogenesis response (ITGA4, ITGAV) were only moderately changed in the infarction at termination, confirming the in vivo PET findings.
Study outline

Conclusions
In this rabbit MI model, we demonstrate the feasibility of monitoring angiogenesis in the healing process non-invasively with PET. The imaging results were confirmed by IHC and gene expression analysis. Moreover, TEE using a dedicated pediatric probe is feasible in the rabbit model, making this a robust and translational medium-sized animal model of myocardial infarction.