The project is agreed as a bilateral collaboration between IBAR and ATOMKI, and the University of Debrecen and IFIN-HH participate in this project voluntarily.
The project is agreed as a bilateral collaboration between IBAR and ATOMKI, and the University of Debrecen and IFIN-HH participate in this project voluntarily. There are two main directions intended by the proposed project: receptor mapping and therapy, using an affibody against the HER2 receptor, combined with a suitable radioisotope. In this sense, the specific objectives are:
Head of Department
Prof. Dr. Stefan Szedlacsek is the Head of the Enzymology Department at the Institute of Biochemistry of the Romanian Academy. He holds a PhD degree in Biotechnology from Polytechnical University of Bucharest as well as a MSc in Organic Synthesis (Polytechnical University, Bucharest) and MSc in Mathematics (University of Bucharest). As a visiting scientist, he performed research in the field of cholesterol metabolism at the University of Illinois at Urbana-Champaign (USA), where he succeeded to evidence a new pathway in the metabolism of oxysterols. He is an “Alexander von Humboldt” fellow and worked in Germany, in the Institute of Biochem More...
Research Assistant
Adina has a Bachelor's degree in Biology and a Master's degree in Applied Genetics and Biotechnology from the Faculty of Biology, University of Bucharest. She is a PhD student and Research Assistant at the Institute of Bio More...
Research Assistant
Andrei-Mihai Vasilescu is a researcher in the Institute of Biochemistry of the Romanian Academy. Andrei-Mihai is currently working in Enzymology in the Enzymology.
Research Assistant
Horea Szedlacsek is a researcher in the Institute of Biochemistry of the Romanian Academy. Horea is currently working in Enzymology in the Enzymology.
Scientific Researcher
Rodica-Aura Badea is a researcher in the Institute of Biochemistry of the Romanian Academy. Rodica-Aura is currently working in Cell Signaling Research in the Enzymology.
Research Assistant
| The Romanian team | The Hungarian team |
|---|---|
|
Dr.Stefan Szedlacsek, Romanian Head of Project Dr.Horea Szedlacsek Dr.Rodica-Aura Badea Asc.Simona Calaras PhD Student Adina-Gabriela Puiu PhD Student Andrei Mihai Vasilescu |
Dr. Szűcs Zoltán , Institute for Nuclear Research (Atomki) Debrecen Dr. Gyula Tircsó, University of Debrecen PhD Student, Károly Brezovcsik, University of Debrecen |
The project comprises three different stages:
I.Preparatory experiments with the main objective being to obtain a stable chemical compound, in turn being subdivided in:
I.1. Smale-scale expression, purification and optimization of the refolding of the anti-HER2 affibody (IBAR).
I.2. Optimized methodology for radioisotope production (ATOMKI & IFIN-HH).
I.3. Comparison among various radiolabeling scenarious using the proposed radioisotopes with chelator agents like: new derivates of DOTA, NODAGA, etc to fit the N-terminal cysteine link to the anti-HER2 affibody (IBAR &ATOMKI, UD)
II.In vivo testing with the objective of testing the behavior of the most promising radiolabeled affibody on cell lines (murine and human kind) This stage subdivides in:
II.1 Large-scale expression, purification and labeling of of anti-HER2 affibody (IBAR)
II.2. Optimizitation of the radiopharmaceutical compounds (IBAR,ATOMKI & IFIN-HH) II.3In vitro radiopharmaceutical testing (IBAR, ATOMKI&IFIN-HH)
III.In vitro testing with the main objective: biodistribution analysis and image quality enhancement using the most promising radiolabeled affibody against HER2 compound (IBAR, ATOMKI &IFN-HH).
The 3,9-PC2ABnpCO2H platform was selected for bioconjugation and for further in vivo studies.
Anti-HER2 affibody was used as a highly specific binding protein, which was expressed in a prokaryotic system, purified by affinity chromatography followed by RP-HPLC.
The specificity of the affibody for HER2 receptors was proved following its conjugation with AlexaFluor555 dye. Finally the affibody was conjugated with the selected 3,9-PC2ABnpCO2H BFC and [52Mn]Mn(II) labelling was performed.
The labelled conjugate was tested in a pilot study by PET imaging using MDA-MB (HER2+) and 4T1 (HER2−) tumour-bearing mice.
The results demonstrate that the [[52Mn]Mn(3,9-PC2ABnpMA)(H2O)]-Cys-HER2-affibody conjugate is an efficient and specific molecular vehicle for targeting and visualizing HER2+ tumours by 52Mn-based PET imaging.
Published papers: Synthesis and characterization of a novel [52Mn]Mn-labelled affibody based radiotracer for HER2+ targeting” Váradi, B., Brezovcsik, K., Garda, Z., Madarasi, E., Szedlacsek, H., Badea, R.-A., Vasilescu, A.-M., Puiu, A.-G., Ionescu, A., Sima, L.-E., Munteanu, C., V.-A., Călăraș, S., Vagner, A., Szikra, D., Toàn, N., M., Nagy, T., Szűcs, Z., Szedlacsek, S., E., Nagy, G., Tircsó, Inorganic Chemistry Frontiers, Accepted: 29 May 2023; First published: 06 Jun 2023; DOI https://doi.org/10.1039/D3QI00356F;
Meeting Abstract: „Labelling anti-HER2-affibodies with Mn-52 via pyclen based bifunctional ligands: from ligand design to in vivo PET/MR experiments”; Varadi, B, ; Garda, Z; Madarasi, E; Brezovcsik, K; Vagner, A; Nagy, T; Garai, I ; Ngo, TM; Puiu, AG ; Vasilescu, AM; Szucs, Z ; Szedlacsek, SE; Nagy, G; Tircso, G at Conference Meeting 35th Annual Meeting of the European-Association-of-Nuclear-Medicine (EANM); Barcelona, Spain; Oct. 15-19, 2022; https://www.webofscience.com/wos/woscc/full-record/WOS:000857046600592
Invited Lecturer in the Fulbright Visiting Scholar Program for the Chemistry Seminar Series at Clarkson University, NY (USA); oral presentations: "Molecular Vehicles for targeted diagnostics and Therapy of Cancer”; 2023 January 20-21;
Romanian Academy