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

Project Information

ImageTumor

Grant agreement ID: 101066639

DOI

10.3030/101066639

Project closed

EC signature date 28 July 2022

Start date 1 June 2023

End date 31 May 2025

Funded under

• Marie Skłodowska-Curie Actions (MSCA)

Total cost

No data

EU contribution

€ 189 687,36

Coordinated by

RUHR-UNIVERSITAET BOCHUM
Germany

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

Reporting period: 2023-06-01 to 2025-05-31

Summary of the context and overall objectives of the project

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.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

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.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

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.