Novel therapies targeting N-WASP protein in breast cancer

Yu, Yannan 2025. Novel therapies targeting N-WASP protein in breast cancer. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Metastasis remains the primary cause of death in breast cancer patients, particularly in triple-negative subtypes, where therapeutic options are limited. This thesis investigates the functional role of neural Wiskott-Aldrich Syndrome protein (N-WASP), a regulator of actin polymerisation and evaluates its potential as a pharmacological target to suppress metastatic behaviour. A series of small-molecule inhibitors were screened, selected and tested, including novel compounds (E06, F02, F04, F06) and clinically approved drugs repurposed for this study (Rucaparib, Aerius, Cyproheptadine). Their effects on breast cancer cell and endothelial cell proliferation, motility, invasion and cytoskeletal architecture were assessed in multiple cell function assays. Among them, F06 consistently showed strong and sustained inhibition of invasive phenotypes. To directly assess N-WASP activity, a pyrene-actin polymerisation bioassay was employed, which confirmed that several compounds, including F06, effectively interfere with N-WASP–mediated actin assembly in vitro. To further explore the downstream consequences of N-WASP inhibition, mass spectrometry-based proteomic and phosphoproteomic analyses were conducted in F06-treated MDA-MB-231 cells. This revealed widespread changes in actin-binding proteins, adhesion-related factors, and signalling intermediates. Network-based analyses (STRING) and pathway enrichment (Reactome) provided insight into the molecular responses to N-WASP disruption, some of which were consistent with those observed in N-WASP knockdown models. Altogether, this work supports the view that N-WASP plays a critical role in promoting breast cancer cell invasiveness and that its inhibition, both by novel and repurposed compounds, has functional and mechanistic effects. These findings provide a basis for future investigation into N-WASP-targeted strategies in metastatic breast cancer.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Schools > Medicine
Date of First Compliant Deposit:	25 November 2025
Last Modified:	02 Dec 2025 09:12
URI:	https://orca.cardiff.ac.uk/id/eprint/182562

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