A Dually Nanobody-Engineered Milk-Derived Extracellular Vesicle Nanomedicine Targeting Tumour-Associated Macrophages and Cancer Cells for Cancer Therapy.

Tumour development and progression are driven by intricate interactions among various cell types within the tumour microenvironment (TME). Targeting a single cell type often fails to eradicate cancer, highlighting the need for strategies to co-target multiple cell types. MicroRNA-21-5p, highly abundant in tumour cells and tumour-associated macrophages (TAMs), exerts strong cancer-promoting effects. Epidermal growth factor receptor (EGFR) is overexpressed in various cancer types, and programmed death-ligand 1 (PD-L1) is expressed predominantly by TAMs in multiple cancers. In this study, we developed a dual-targeted engineered milk-derived extracellular vesicles system (7D12/KN035-iEVs), decorated with 7D12 (an EGFR nanobody) and KN035 (a PD-L1 nanobody), to specifically deliver miR-21-5p inhibitors into EGFR and/or PD-L1 tumour cells and TAMs, thereby inhibiting tumour progression while reprogramming immunosuppressive TME. Notably, this dual-targeting nanomedicine synergistically inhibits tumour growth when combined with immunotherapy and radiotherapy. In summary, this mEV-based nanomedicine represents a promising universal strategy for cancer treatment, offering a versatile platform for targeting multiple components of the TME.