Periodic Reporting for period 1 - NO-CANCER-NET (Advanced Engineering of Nitric Oxide Based Therapeutics for Triple Negative Breast Cancer Training Network)
Okres sprawozdawczy: 2023-10-01 do 2025-09-30
NO-CANCER-NET (Advanced Engineering of Nitric Oxide Based Therapeutics for Triple Negative Breast Cancer Training Network) aims to set a landmark in the present European Higher Education Area. Our training initiative uses molecular epidemiological informed approaches for the identification of molecular targets, development of companion diagnostics and a panel of novel therapeutic approaches, which addresses an unmet medical need for triple negative breast cancer (TNBC). The NO-CANCER-NET Doctoral Network (DN) addresses two fundamental gaps in European training: 1) a network where a core of Doctoral Candidates (DCs) can develop and share a basic understanding of molecular target discovery, models of tumour biology assessment, drug synthesis and delivery, developing diagnostics, handling complex spatial imaging and omics expression data profiles; and 2) solid interdisciplinary training in experimental, computational and clinical research. NO-CANCER-NET will equip our DCs with the required knowledge and training to understand the complexities of bench to bedside cancer research, the importance of an interdisciplinary approach and an understanding of the fundamentals of related disciplines. This will be of great importance in preparing DCs to work in academia, industry and clinical environments. To achieve this training, the individual DC projects and work packages (WPs) will be integrated and have interdependency. Beneficiaries and associated partners are already collaborating in an existing International Society for Nitric Oxide and Cancer (www.isnoc.org) covering academic, clinical, and industrial sectors from 7 countries.
The overall aim of the proposed research is to explore the potential of therapeutics targeting NO and NO-regulated pathways in TNBC. We define 3 WPs which will address 4 main research objectives (RO).
WP1 – Biomarker discovery for effective patient stratification of inflammation-related TNBC.
RO1 - To identify the key biomarkers and signalling pathways indicative of NO driven TNBC. There is currently no reliable way to predict treatment response or stratify TNBC patients for treatment. WP1 will facilitate the development of biomarkers allowing an optimal patient stratification and identification of the appropriate NO related treatment choice.
WP2 - NO-based drug development and delivery
RO2 - To develop a panel of novel therapeutics that target NO and NO regulated pathways for the improved treatment of TNBC.
RO3 - To generate appropriate drug delivery systems for optimal stable delivery of NO related therapeutics and for appropriate combination with existing treatments. There is a lack of an effective, safe and targeted approach to deliver appropriate amounts of NO alone or in combination with other therapeutic approaches at target tissues. In WP2 this technical and medical gap will be addressed to reach the above objectives. Additionally, targeted therapies do not yet exist for TNBC patients. WP2 will exploit associations between NOS signalling and TNBC patient survival to develop and deliver novel NO-targeted therapies.
WP3 - 3D tumour models to investigate tumour biology and drug response
RO4 - To develop 3D models of TNBC that reflect different NO states, to allow for improved understanding of NO biology and testing of novel therapeutics.
The effects of NO in TNBC disease processes are complex, dependent on NO concentration and tumour genotype. WP3 will develop 3D models to enhance our understanding of the complexities of NO biology in TNBC.
The overall aim of the proposed research is to explore the potential of therapeutics targeting NO and NO-regulated pathways in TNBC. We define 3 WPs which will address 4 main research objectives (RO).
WP1 – Biomarker discovery for effective patient stratification of inflammation-related TNBC.
RO1 - To identify the key biomarkers and signalling pathways indicative of NO driven TNBC. There is currently no reliable way to predict treatment response or stratify TNBC patients for treatment. WP1 will facilitate the development of biomarkers allowing an optimal patient stratification and identification of the appropriate NO related treatment choice.
WP2 - NO-based drug development and delivery
RO2 - To develop a panel of novel therapeutics that target NO and NO regulated pathways for the improved treatment of TNBC.
RO3 - To generate appropriate drug delivery systems for optimal stable delivery of NO related therapeutics and for appropriate combination with existing treatments. There is a lack of an effective, safe and targeted approach to deliver appropriate amounts of NO alone or in combination with other therapeutic approaches at target tissues. In WP2 this technical and medical gap will be addressed to reach the above objectives. Additionally, targeted therapies do not yet exist for TNBC patients. WP2 will exploit associations between NOS signalling and TNBC patient survival to develop and deliver novel NO-targeted therapies.
WP3 - 3D tumour models to investigate tumour biology and drug response
RO4 - To develop 3D models of TNBC that reflect different NO states, to allow for improved understanding of NO biology and testing of novel therapeutics.
The effects of NO in TNBC disease processes are complex, dependent on NO concentration and tumour genotype. WP3 will develop 3D models to enhance our understanding of the complexities of NO biology in TNBC.
Progress for WP1 & RO1
WP1’s principal aim is to deliver biological insights for NO/S-nitrosylation-based TNBC stratification, enabling the identification of patient subgroups with distinct inflammatory and metabolic profiles. This knowledge will be fundamental for the development and application of NO-related therapeutic strategies (WP2) and guide the design of physiologically relevant 3D and in vivo models (WP3).
WP1 brings together three complementary doctoral projects that collectively span mechanistic, translational, and systems-level analyses. All three doctoral projects related to WP 1 and RO 1 (DC1, DC2 & DC3 based at DCS, TCD & UL respectively) have completed their first year of their PhD studies and are making significant progress.
• DCS: GSNOR as a new diagnostic marker of cancer and inflammatory diseases (DC1)
• TCD: Inflammatory caspases as regulators of NO levels during triple negative breast cancer (DC2)
• UL: Spatial modelling of cancer metabolic networks on multi-cell population level (DC3)
Experimental studies to date are focusing on:
1. Identifying S-nitrosylated proteins and metabolic alterations associated with impaired denitrosylation.
2. Defining inflammatory caspases as modulators of NO production in tumour and stromal compartments
3. Validating candidate biomarkers in clinically relevant models.
These efforts are tightly integrated with advanced computational approaches, including bulk, single-cell, and spatial transcriptomics coupled to genome-scale metabolic network modelling, to reconstruct NO-driven metabolic states.
Progress for WP2 and RO2/RO3
WP 2 comprises four complementary projects that converge on a common objective: developing and validating nitric oxide (NO)-centred therapeutic strategies to address the clinical challenges of triple-negative breast cancer (TNBC). All four doctoral projects related to WP2 and RO2/RO3 (DC4, DC5, DC6 & DC7 based at EPHE-PSL, ISOF-CNR, UNIUD & GALWAY respectively) are also making significant progress.
• EPHE-PSL: Sequential chemo – NO donor approaches leveraging inflammation-related TNF-family signalling (DC4)
• ISOF-CNR Tumour-microenvironment–responsive multimodal conjugates that combine NO/ROS generation with chemotherapeutic payloads (DC5)
• UNIUD In vitro validation of photoactivable NO and ROS-releasing molecules for photodynamic therapy (DC6)
• GALWAY: Biomaterial-enabled NO modulation through hydrogel delivery of NO-scavenging therapeutics, supported by the development of dedicated real-time electrochemical NO-detection platforms (DC7)
Progress for WP3 and RO4
WP 3 aims to develop 3D models of TNBC that reflect different NO states to allow for improved understanding of NO biology and for testing of novel therapeutics. The experimental work includes the set-up and optimisation of 3D TME models in relation to clinical features and to allow the study of separate cellular components of the TME; the assessment of NO and REDOX signaling in TME crosstalk; the characterization and selection of NO-based therapeutics acting in TME crosstalk and the functional responses to established conventional antitumor drugs. All three doctoral projects below related to WP3 and RO4 (DC8, DC9 & DC10 based at US, UNISI & GALWAY respectively) are making significant progress.
• US: Impact of Atezolizumab+Paclitaxel, and tyrosine kinase inhibitors or Atezolizumab +Bevacizumab in a 3D culture constituted by TNBC and fibroblast, or SNU449 and stellate cells, respectively. It was assessed the impact of NO inhibitors or donors (DC 8)
• UNISI: Tumour-endothelial cell interplay under NO based drugs (DC 9)
• GALWAY: Developed 3D spheroid models of iNOS cancer biology in Triple Negative Breast Cancer and examined impact on chemotherapeutic response (DC 10)
WP1’s principal aim is to deliver biological insights for NO/S-nitrosylation-based TNBC stratification, enabling the identification of patient subgroups with distinct inflammatory and metabolic profiles. This knowledge will be fundamental for the development and application of NO-related therapeutic strategies (WP2) and guide the design of physiologically relevant 3D and in vivo models (WP3).
WP1 brings together three complementary doctoral projects that collectively span mechanistic, translational, and systems-level analyses. All three doctoral projects related to WP 1 and RO 1 (DC1, DC2 & DC3 based at DCS, TCD & UL respectively) have completed their first year of their PhD studies and are making significant progress.
• DCS: GSNOR as a new diagnostic marker of cancer and inflammatory diseases (DC1)
• TCD: Inflammatory caspases as regulators of NO levels during triple negative breast cancer (DC2)
• UL: Spatial modelling of cancer metabolic networks on multi-cell population level (DC3)
Experimental studies to date are focusing on:
1. Identifying S-nitrosylated proteins and metabolic alterations associated with impaired denitrosylation.
2. Defining inflammatory caspases as modulators of NO production in tumour and stromal compartments
3. Validating candidate biomarkers in clinically relevant models.
These efforts are tightly integrated with advanced computational approaches, including bulk, single-cell, and spatial transcriptomics coupled to genome-scale metabolic network modelling, to reconstruct NO-driven metabolic states.
Progress for WP2 and RO2/RO3
WP 2 comprises four complementary projects that converge on a common objective: developing and validating nitric oxide (NO)-centred therapeutic strategies to address the clinical challenges of triple-negative breast cancer (TNBC). All four doctoral projects related to WP2 and RO2/RO3 (DC4, DC5, DC6 & DC7 based at EPHE-PSL, ISOF-CNR, UNIUD & GALWAY respectively) are also making significant progress.
• EPHE-PSL: Sequential chemo – NO donor approaches leveraging inflammation-related TNF-family signalling (DC4)
• ISOF-CNR Tumour-microenvironment–responsive multimodal conjugates that combine NO/ROS generation with chemotherapeutic payloads (DC5)
• UNIUD In vitro validation of photoactivable NO and ROS-releasing molecules for photodynamic therapy (DC6)
• GALWAY: Biomaterial-enabled NO modulation through hydrogel delivery of NO-scavenging therapeutics, supported by the development of dedicated real-time electrochemical NO-detection platforms (DC7)
Progress for WP3 and RO4
WP 3 aims to develop 3D models of TNBC that reflect different NO states to allow for improved understanding of NO biology and for testing of novel therapeutics. The experimental work includes the set-up and optimisation of 3D TME models in relation to clinical features and to allow the study of separate cellular components of the TME; the assessment of NO and REDOX signaling in TME crosstalk; the characterization and selection of NO-based therapeutics acting in TME crosstalk and the functional responses to established conventional antitumor drugs. All three doctoral projects below related to WP3 and RO4 (DC8, DC9 & DC10 based at US, UNISI & GALWAY respectively) are making significant progress.
• US: Impact of Atezolizumab+Paclitaxel, and tyrosine kinase inhibitors or Atezolizumab +Bevacizumab in a 3D culture constituted by TNBC and fibroblast, or SNU449 and stellate cells, respectively. It was assessed the impact of NO inhibitors or donors (DC 8)
• UNISI: Tumour-endothelial cell interplay under NO based drugs (DC 9)
• GALWAY: Developed 3D spheroid models of iNOS cancer biology in Triple Negative Breast Cancer and examined impact on chemotherapeutic response (DC 10)
Our DCs have completed the 1st year of their PhD studies. At this point it is premature to report on results beyond the state of the art,