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)
