Membrane-anchored tissue inhibitor of matrix metalloproteinase (TIMP)-1 promotes cell death in head and neck cancer by inducing DNA damage, accumulating collagen II and disrupting cell survival mechanisms

Head and neck squamous cell carcinoma (HNSCC) remains a significant global health challenge with limited therapeutic options. Here, we investigate a novel membrane-anchored variant of Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1), designated T1^Pr, as a potential therapeutic agent against HNSCC. Utilizing a comprehensive approach involving biochemical, cellular, animal, mRNA-sequencing, and omics techniques, we demonstrate that T1^Pr exhibits remarkable anti-tumorigenic effects in CAL27 cells in vitro and in vivo. Phospho-proteomics analysis revealed that T1^Pr induces profound changes in the cellular phosphorylation landscape, particularly targeting critical cell cycle regulators like CDK1 and CSNK2A1. Further, T1^Pr disrupts nuclear transport and cell cycle pathways, causes chromosomal DNA damage possibly by impeding the nuclear localization of membrane-associated MMPs and ADAM proteinases. Notably, T1^Pr triggers significant extracellular matrix remodeling, including collagen II accumulation, and suppresses numerous pro-tumor genes and proteins. T1^Pr also upregulates tumor-suppressive genes like PAWR, TPM1, and THBS1 linked to cytoskeletal stability and apoptosis induction. Functionally, T1^Pr drives CAL27 cells toward apoptosis through persistent DNA damage and disrupted mitotic checkpoints. Importantly, T1^Pr does not compromise immune checkpoint mechanisms. These findings position T1^Pr as a promising molecular therapeutic approach for targeting invasive cancers, particularly HNSCC, by orchestrating a comprehensive cellular shutdown through multiple interconnected mechanisms.