Periodic Reporting for period 2 - NEOSETAC (New Selenium-based Targeted Nanocapsules to treat Breast Cancer)
Reporting period: 2020-01-01 to 2023-10-31
In the NEOSETAC project, breast cancer will be the focus of study being the most common cancers in women. It comprises of 10.4% of all the cancer incidences among women, causing 411.093 deaths per year all over the world. In Europe, there is more than twice of the amount of new breast cancer cases annually than new cases of cancer in any other place.
Treatment for breast cancer is dependent on disease stage, histologic and molecular subtypes and menopausal status. Further aspects influencing treatment choice for early breast cancer include balancing the risk of relapse with the benefit of intervention and patient factors such as the impact of treatment on fertility. Surgery (mastectomy or breast-conserving surgery with or without lymph node dissection) and radiotherapy play an important role in early breast cancer: systemic therapy may be used for almost all women and is the predominant treatment for those with advanced disease
The NEOSETAC project will demonstrate the anti-cancer therapeutic potential of a novel Selenium-based therapy for the management of breast cancer. A recent clinical study shows that high selenium exposure could decrease cancer risk, especially high plasma/serum selenium and toenail selenium. NEOSETAC will demonstrate the potential of a Selenium-based compounds cocktail to attack breast cancer cells; this compounds will be encapsulated in a targeted nanocapsule that will drive the compounds directly to the cancer cells. The overall research and development goal of the proposed action is twofold: to design and manufacture the Se-nanocapsules to treat breast cancer and demonstrate their therapeutic potential in pre-clinical studies.
Treatment for breast cancer is dependent on disease stage, histologic and molecular subtypes and menopausal status. Further aspects influencing treatment choice for early breast cancer include balancing the risk of relapse with the benefit of intervention and patient factors such as the impact of treatment on fertility. Surgery (mastectomy or breast-conserving surgery with or without lymph node dissection) and radiotherapy play an important role in early breast cancer: systemic therapy may be used for almost all women and is the predominant treatment for those with advanced disease
The NEOSETAC project will demonstrate the anti-cancer therapeutic potential of a novel Selenium-based therapy for the management of breast cancer. A recent clinical study shows that high selenium exposure could decrease cancer risk, especially high plasma/serum selenium and toenail selenium. NEOSETAC will demonstrate the potential of a Selenium-based compounds cocktail to attack breast cancer cells; this compounds will be encapsulated in a targeted nanocapsule that will drive the compounds directly to the cancer cells. The overall research and development goal of the proposed action is twofold: to design and manufacture the Se-nanocapsules to treat breast cancer and demonstrate their therapeutic potential in pre-clinical studies.
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.
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.
In NEOSETAC, the main innovative aspect of the new therapy is the therapeutic cocktail of selenium compounds used in the formulation that improves the antitumor potency. In previous studies, researchers had used different selenium compounds separately to treat different types of cancer [69-71], but NEOSETAC proposes a mixture of these compounds specifically designed to improve the effect of the drug. Moreover, their effectiveness will be improved by the active targeting, coupling them with specific antibodies.
NEOSETAC is expected to have substantially less side effect, thus increasing the quality of life of patients during second line therapy. As it is a less strenuous treatment, NEOSETAC will have an impact on both the survival as well as the quality of life in this patient population.
Besides quality of life, substantial economic impacts are to be expected from NEOSETAC. This is an important attribute for the sustainability of health care systems in an age where health care budgets are increasingly constraint; thus, this is a compelling reason to improve the research within worldwide.
NEOSETAC represents an exceptional and unique opportunity for both ER and ESR to enhance their know-how through the scheduled secondments between academy and industry to put a practical emphasis on the subject of their research activities, as they will be directly present in the industry. Furthermore significant contribution to knowledge and technology transfer is expected from these academic and companies.
NEOSETAC is expected to have substantially less side effect, thus increasing the quality of life of patients during second line therapy. As it is a less strenuous treatment, NEOSETAC will have an impact on both the survival as well as the quality of life in this patient population.
Besides quality of life, substantial economic impacts are to be expected from NEOSETAC. This is an important attribute for the sustainability of health care systems in an age where health care budgets are increasingly constraint; thus, this is a compelling reason to improve the research within worldwide.
NEOSETAC represents an exceptional and unique opportunity for both ER and ESR to enhance their know-how through the scheduled secondments between academy and industry to put a practical emphasis on the subject of their research activities, as they will be directly present in the industry. Furthermore significant contribution to knowledge and technology transfer is expected from these academic and companies.