The general scientific progress of the action during the period covered by this report are:
WP1. The composition of the seleno-compounds cocktail has been achieved. Citotoxicity of different variations (Individual Se compounds and Se cocktail) have been examined in breast cancer cell lines (mouse breast cancer cell line 4T1, and human breast cancer cell lines MDA and MCF-7) and control cell lines 3T3). Design and characterization of the different nanosystems loaded with Se-cocktail has been started. A database of cancer treatments (dose, timing, etc) from real cancer patients was set up in Sweden to facilitate further studies of the drugs and the optimal doses.
WP2. Three possible breast cancer cell markers were selected upon the bibliography research done: Epithelial cell adhesion molecule (EpCAM); Epidermal Growth Factor Receptor (EGFR); and Homing cell adhesion molecule (HCAM or CD44). The antibodies against these markers were selected based on their reactivity (mouse and human) and their intended application (flow cytometry-FACS and immunofluorescence-IF).
WP3. A compilation of the data obtained in WP1 and WP2 has been done.
WP4. The work package will start on month 32.
WP5. Dissemination and Communication Strategy implemented and monitored to exploit the potential of long-lasting cooperation among the involved partners as well as to create new networks to consolidate and widen the existing ones among the academic and industrial communities involved.
WP6 The implemented management framework has been updated to keep ensuring the achievement of project aims within the time and budget and ensures communication and coordination among the partners and with the REA, including the employment of a quality assurance procedure to check project performance, impact, risks and visibility. Fluent communication with the PO to translate project progress, status of secondments and implement the requested amendments due to the inclusion of a new beneficiary. Further information available through D6.1
WP7 A copy of project authorization, covering also the work with genetically-modified animals has been obtained and D7.1 submitted.
Regarding the main results achieved:
The organic selenium compound, p-XSC, with highest anti-cancer potency was selected to further investigate the synergistic effect with other selelium compounds and/or other chemotherapeutic drugs. A strong synergistic effect on breast cancer cells was observed when p-XSC used as adjuvant treatment besides clinically established drugs, i.e. Gemcitabine (GEM) and Doxorubin (DOX).
Therefore, p-XSC/GEM and p-XSC/DOX were selected as the cocktail composition to be loaded into the nanocarrier system.
In vitro cytotoxicity investigations were carried out using different breast cancer cell lines including mouse breast cancer cell line 4T1, human breast cancer cell lines: MCF7 and MDA-MB-231, and control cell line, mouse fibroblast cell line 3T3.
Both p-XSC/GEM and p-XSC/DOX cocktail showed a strong synergistic effect on breast cancer cell lines: 4T1, human breast cancer cell lines: MCF7 and MDA-MB-231, while such a synergistic effect was not observed on fibroblast cell line. Thus, in the present of selenium compound, the dose of GEM and DOX can be reduced, therefore the adverse effects and/or the toxicity caused by using high doses of GEM and DOX could also be reduced.
Besides the indirect analysis method LC-MS to quantify selenium compounds, we also used for direct analysis X-ray absorption spectroscopy (XAS) to further confirm albumin/plasma protein binding and Se-S bond. XAS features minimal sample preparation prior to data acquisition compared to hyphenated techniques including LC-MS. Therefore, it has less effect on species transformation and could provide atomic information of Se in complex matrix. XAS analysis validated the covalent Se-S binding model and demonstrated notable albumin binding of selenium compounds in context of plasma.
