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Theranostics 2019; 9(5):1474-1489. doi:10.7150/thno.29255 This issue Cite

Research Paper

Molecular Imaging of Factor XIII Activity for the Early Detection of Mouse Coronary Microvascular Disease

Zhen W. Zhuang^1,2,3✉, Yang Huang³, Rong Ju⁴, Mark W. Maxfield⁵, Yongming Ren³, Xiangning Wang^1,2, Xinlu Wang^1,2, Mitchel R. Stacy^1,2, John Hwa^1,3

1. Section of Cardiovascular Medicine, Yale University School of Medicine, New Haven, CT, USA.
2. Yale Translational Research Imaging Center, Dept. of Internal Medicine.
3. Yale Cardiovascular Research Center, Dept. of Internal Medicine, New Haven, CT 06510-3221, USA.
4. State key laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China.
5. Dept. of Surgery, Yale University School of Medicine, New Haven, CT, USA.

✉ Corresponding author: Zhen W. Zhuang, MD, Section of Cardiovascular Medicine, Yale University School of Medicine, 375 Congress Ave, LSOG 120C, New Haven, CT 06510. Tel: 203-231-9084; Fax: 203-737-6236; e-mail: zwz2yale.edu

Abstract

Coronary microvascular disease (MVD) remains a major clinical problem due to limited mechanistic understanding and a challenging diagnosis. In the present study we evaluated the utility of targeted imaging of active factor XIII (FXIII) for detection of coronary MVD associated with thrombus. We hypothesized that a high specificity and sensitivity FXIII targeted radiolabeled probe can serve as a biomarker for cross-linked thrombi in the microvasculature, and thus an indicator for underlying coronary MVD. To evaluate this approach, a coronary MVD model was established for local induction of singlet oxygen and reactive oxygen species (ROS) via a photochemical reaction (PCR).

Methods: PCR was used to induce endothelial injury and microthrombi via focal over-production of ROS only in the coronary microvasculature. Oxidative stress was initially evaluated in primary coronary endothelial cells to optimize parameters of PCR, which were then translated to in vivo experiments. To develop the coronary MVD model, 64 mice were assigned to one of four groups after thoracotomy: 1) sham control; 2) rose bengal; 3) green light; or 4) their combination. Following interventions, the mice underwent transmission electron microscopy, fluorescent myocardial perfusion, coronary angiography, and immunohistochemical staining. Echocardiography (n = 12) and gene expression (n = 10) studies were also performed after MVD induction to monitor serial changes in cardiac function and explore possible mechanisms. To diagnose early onset MVD, FXIII radioactivity was assessed in 104 mice using ex vivo gamma well counting (GWC) and in 14 mice using in vivo serial single photon emission computed tomography / computed tomography (SPECT/CT) imaging of a FXIII targeted technetium-labeled probe (^99mTc-NC100668).

Results: In vitro experiments demonstrated that photosensitizer concentration and light illumination time were critical parameters for PCR. In vivo experiments demonstrated manifestations of clinical MVD, including endothelial damage, a “no flow zone,” arteriole rarefaction with patent epicardial coronary arteries, infiltration of inflammatory cells in the PCR-treated region, and preserved cardiac function. Gene expression also demonstrated a pro-thrombotic and impaired fibrinolytic status. In the early stages of MVD, enhanced FXIII activity was confirmed within the MVD region using GWC and in vivo SPECT/CT imaging.

Conclusion: Our results demonstrate that molecular imaging of FXIII activity may allow for early detection of coronary MVD associated with thrombus, in a novel pre-clinical model.

Keywords: coronary microvascular disease, factor XIII, photochemical reaction, micro-computed tomography, single photon emission computed tomography

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