Progress is according to plan. From over 200 candidates, we recruited 11 DCs, employed full-time, and enrolled in a PhD programme. All governing boards (supervisory board, DC-board, data management and intellectual property board, educational board) have been constituted and work well. Beside the regular online journal clubs and scientific meetings, the DCs participated in two eRaDicate schools (in person in Santiago de Compostela and one online). They presented their work receiving feedback on their presentation skills and scientific projects. In progress outreach includes social media engagement (https://www.linkedin.com/company /eradicateprojecteu/), the eRaDicate website (
https://www.eradicate-project.eu/(se abrirá en una nueva ventana)) and participation in public science.
In the scientific work packages (WP) we address mechanistic aspects of RARs and VDR mainly in leukaemia (WP1); focus on solid tumours (WP2); on chemistry and pharmacy (WP3).
We have shown that expression of the RARG gene in human bone marrow samples from acute myeloid leukaemia (AML) patients is similar to that in healthy persons (DC-UWr2). A combination of vitamin D analogues and electrophilic agents had synergistically reduced growth of AML cells (DC-BGU).
DC-MUV1 has determined the effect of RAR- and VDR-targeting compounds on the proliferation and migration of patient-derived primary high-grade serous ovarian cancer (HGSOC) cells, and commercially available colorectal cancer (CRC) cell lines. The target compounds have been tested individually, in combination with each other, and with existing chemotherapeutics using live-cell cytometry. DC-MUV1 quantified the expression of the nuclear receptors and the cancer stem cell markers CD44 and CD133 at mRNA and protein level in selected HGSOC cell models. DC-MUV2 focused on three prostate cancer (PCa) 3D models obtained from genetically modified mice. All organoids expressed the RARγ and responded in a dose-dependent manner to treatment with the RAR antagonist AGN205728 and the vitamin D agonist PRI-5202, supporting the robustness of the models regarding validation of the therapeutic efficacy of our compounds before moving to in vivo studies. DC-USC developed both 2D and 3D models for triple-negative breast cancer cells and has optimized these for drug treatment studies, with viability and apoptosis, using MTT assays and confocal immunofluorescence.
DC-UoW has been working on the structural and biophysical characterisation of full-length hVDR, its ligand-binding domain (LBD), and its interactions with ligands. To express the full-length hVDR and VDR-LBD in mammalian systems, DC-UoW tested multiple constructs, promoters, affinity tags, and host cell lines to optimise yield and solubility. Using computational molecular docking and modelling, predicted binding modes and refined ligand selection.
To develop a dual-function RARγ/VDR hybrid molecule, DC-MUW established a comprehensive in silico design, selected a lead candidate from 14 prototype structures and designed a convergent 14-step synthetic route based on three modular building blocks. In parallel, DC-MUW has synthetised novel VDR agonists with improved metabolic stability that outperformed 1,25-dihydroxyvitamin D₃. The reference RARγ antagonist, AGN205728, was fully characterised, verified, and distributed to DCs for biological evaluation.
DC-TCD focused on establishing foundational and systematic pre-formulation strategies for vitamin D analogues using cholecalciferol and ergocalciferol as model compounds. Amorphous solid dispersions (ASDs) were successfully prepared at high drug loading using industry-relevant processing methods, including spray drying, solvent evaporation, and solvent-free mechanochemical ball milling with selected polymers. Physicochemical characterisation confirmed the formation of fully amorphous systems showing improved solubility and provided insight into stability, miscibility, and key processing–property relationships.
