X-Ray Fluorescence Imaging of Metal Complexes in Hypoxic Tumor Spheroids

Informazioni relative al progetto

ImageTumor

ID dell’accordo di sovvenzione: 101066639

DOI

10.3030/101066639

Progetto chiuso

Data della firma CE 28 Luglio 2022

Data di avvio 1 Giugno 2023

Data di completamento 31 Maggio 2025

Finanziato da

• Marie Skłodowska-Curie Actions (MSCA)

Costo totale

Nessun dato

Contributo UE

€ 189 687,36

Coordinato da

RUHR-UNIVERSITAET BOCHUM
Germany

Periodic Reporting for period 1 - ImageTumor (X-Ray Fluorescence Imaging of Metal Complexes in Hypoxic Tumor Spheroids)

Periodo di rendicontazione: 2023-06-01 al 2025-05-31

Sintesi del contesto e degli obiettivi generali del progetto

Context
According to estimates from the World Health Organization (WHO) in 2020, cancer is the second leading cause of death with approximately 10 million deaths. The field of metal-based anticancer drugs has been on the rise ever since the discovery of the anticancer properties of cisplatin in 1965, which is in daily clinical use against various types of cancer. Many compounds failed to enter clinical studies since their anti-cancer activity was mainly studied in two-dimensional (2D) monolayer cells which does not mimic the complex tumor microenvironment in humans. In the human body the cancer tumours grow in 3-dimensional structure, hence it is vital to study the anticancer activity of a drug molecules in the 3D tumours rather in 2D mono layer cells. We have investigated the structure activity relationship of Rhenium-metal complexes to realise the effect of the cytotoxicity in 3D tumor spheroids.
Key objectives of the project
1. Synthesis and characterization of Re-metal complexes.
2. Study the cytotoxicity in 2D and 3D tumor spheroids and compare and investigate the cell death mechanism.
3. Investigate the cellular uptake and localization in cancer cells with confocal fluorescence microscopy where applicable.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

The activities performed and the main achievements

-Successfully synthesized Rhenium tricarbonyl complexes and characterized using 1HNMR 13CNMR, ESI and IR spectroscopy methods.
- Photophysical properties were measured by UV-Vis, and fluorescence spectroscopy in acetonitrile.
-Firstly, anticancer activity was tested by MTT assay in 2 dimensional (2D) Hela cells for 12 organometallic Re-complexes and then ruled out the non-cytotoxic compounds. Next, further MTT assay studies were conducted in 2D and cell titerglo assay in 3D tumour spheroids with only four compounds that showed interesting cytotoxic activity in the other three cell lines namely HeLa, MCF-7 and GM5756 fibroblast cell lines at normoxic conditions.
-Cellular uptake quantification of metal was investigated in HeLa cells using inductively coupled mass spectrometry at 37 °C, 10 % CO2
-Cellular localisation of these complexes were investigated using confocal microscopy
-Partition coefficient measurement/ Lipophilicity measurements were conducted using octanol/water system.
-The possibility of generation of reactive oxygen species (ROS) was detected by using DCFDA studies using microwell plate reader.
-Manuscript preparation is under progress and results will be communicated for publication.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

We have designed and synthesized the 11 Re-tricarbonyl complexes and investigated the structure activity relationship of complexes. The research results obtained in this project were interesting in terms of understanding the anticancer activity of Re-metal complexes in three-dimensional tumor spheroids and still further research is needed to investigate the metal complex penetration into the tumor spheroids. The metal complexes investigated in this project were non- fluorescent in nature and was not easy to detect them using classical confocal microscopy in 3D tumor spheroids. Hence, in the future we are aiming to detect this metal complex presence in solid tumors using the X-ray fluorescence imaging methods. The results of this project do not have potential to access the markets and finance or commercialization.