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Varga Zoltán
In vivo Biodistribution of Extracellular Vesicles: Developing Efficient Radiolabeling Techniques

Aug 29 - kedd

14:00 – 14:20

Orvosi biofizika és sugárbiológia

E14

In vivo Biodistribution of Extracellular Vesicles: Developing Efficient Radiolabeling Techniques

Zoltán Varga1, Kinga Ilyés1, Dávid Szöllősi2, Ildikó Horváth2, Domokos Máthé2,3, Krisztina Németh4,5, Viola Tamási4,*, Edit I Buzás4,5,6, Krisztián Szigeti2

1 Biological Nanochemistry Research Group, Research Centre for Natural Sciences

2 Department of Biophysics and Radiation Biology, Semmelweis University

3 HCMM-SE In Vivo Imaging Advanced Core Facility

4 Department of Genetics, Cell- and Immunobiology, Semmelweis University

5 ELKH-SE Translational Extracellular Vesicle Research Group

6 HCMM-SE Extracellular Vesicle Research Group

*current affiliation: Department of Molecular Biology, Semmelweis University

The understanding of extracellular vesicle (EV) biodistribution plays an important role in advancing circulating biomarker research. Nuclear imaging techniques like single-photon emission computed tomography (SPECT) hold potential, but the literature on radioisotope labeling of EVs for in vivo studies remains scarce. This presentation explores the evolution of novel radiolabeling methods, focusing on the development and comparative evaluation of various Tc99m radiolabeling strategies.

Our initial method involved the radioisotope labeling of erythrocyte-derived EVs using the Tc99m-tricarbonyl complex [1]. In vivo SPECT/CT biodistribution studies in mice showed that intravenously administered Tc99m-labeled EVs primarily accumulated in the liver and spleen. Our observations suggested good in vivo stability, with a minor fraction of the radioactive label detaching from the EVs. Next, we explored an alternative approach using Tc99m-HYNIC-Duramycin to label cell-derived EVs [2]. Duramycin, a membrane-active peptide, specifically labels EVs, resulting in higher labeling efficiency. Following previous observations, significant uptake of EVs in the liver and the spleen was observed.

The latest experimental focus is on the use of recombinant proteins with His-tag in conjunction with the Tc99m-tricarbonyl complex to label EVs. Preliminary investigations indicate promising superior performance with this method compared to the previous techniques.

This presentation will provide an in-depth comparison of these methods, emphasizing their development process and potential implications for advancing in vivo EV imaging studies. Through a critical evaluation of their advantages and potential limitations, we aim to foster a greater understanding of efficient tracking of EV biodistribution for future research.

References

[1] Varga Z et al. (2016) Cancer Biother. Radiopharm. 31: 168-173.

[2] Németh K et al. (2021) Cell Mol Life Sci 78: 7589–7604