Unravelling the in vivo traits of vasculogenic mimicry.

UNLABELLED: Vasculogenic mimicry describes the ability of cancer cells to acquire endothelial properties, forming vessel-like channels that facilitate tumour blood supply and can drive resistance to anti-angiogenic therapies. While the molecular drivers have been well-explored in cell cultures and biopsies, vasculogenic mimicry remains a clinical blind spot as no methods exist to detect and monitor such vascular networks non-invasively in vivo. METHODS: Here, we introduce a "topology-first" framework to define vasculogenic mimicry biomarkers and elucidate its spatiotemporal dynamics using label-free optical and photoacoustic imaging across scales. Using graph theory analysis, we decoded the unique vascular architecture of 4T1 and 4T1-T murine breast cancer models in response to anti-angiogenic treatment with Axitinib. RESULTS: We found stable meshed networks in vitro and blood-containing circular structures in vivo serve as a key topological fingerprint for vasculogenic mimicry, conserved also in human breast cancer models. Crucially, these structures remained remodelling-resistant under anti-vascular therapy, signalling a potential functional mechanism of therapeutic escape. CONCLUSIONS: Our results provide the first evidence that vasculogenic mimicry features can be mapped non-invasively through topology, offering a potential future companion diagnostic and method to guide the development of next-generation, multi-target anti-vascular therapeutics.