Early detection of cancer remains one of the most urgent challenges facing European healthcare systems. Across the EU, cancer causes 1.3 million deaths annually, and prostate and colorectal cancers remain among the most prevalent and difficult to diagnose at early stages. Current diagnostic tools rely predominantly on protein expression levels or genetic markers, which often fail to reflect the functional biochemical activity driving disease progression. One such activity is that of serine proteases- enzymes whose dysregulation contributes to cancer invasion, metastasis, and altered cell–microenvironment interactions. Among these, TMPRSS2 has emerged as a clinically relevant but poorly understood protease, implicated in prostate cancer biology and several epithelial cancers. Despite its relevance, there is a lack of sensitive and selective tools to detect its active form in cells, tissues, or patient-derived samples.
This gap- between the biological importance of TMPRSS2 and the limited availability of chemical tools to study it- formed the central motivation for the MULTIGLOW project. The project set out to design, synthesize, and validate a new generation of multi-modal, chemiluminescent and activity-based probes capable of selectively reporting TMPRSS2 activity in vitro, in cell systems, and in clinically relevant samples. By focusing on enzyme activity rather than expression alone, the project aligns strongly with the EU’s scientific and societal priorities, including Europe’s Beating Cancer Plan, the mission for cancer prevention and early detection, and the objective of building a more resilient healthcare system under the “Economy That Works for People” policy area.
The project pursued four interconnected scientific goals:
1) Development of innovative chemical probes – including chemiluminescent substrates, classical activity-based probes, and multi-modal (luminescent + fluorescent/inhibitory) tools selective for TMPRSS2 and other P1-arginine serine proteases.
2) Biochemical characterization and selectivity profiling – validating probe performance and identifying potent, selective lead candidates for biological applications.
3) Application of probes in biological systems – using the most promising tools to detect TMPRSS2 activity in cancer cell lines, 3D spheroids, and in complex samples, demonstrating their diagnostic and imaging potential.
4) Initial assessment of clinical relevance – examining whether TMPRSS2 activity can be detected in patient-derived biological fluids, evaluating feasibility, limitations, and future diagnostic pathways.
Together, these objectives form a pipeline from molecular design to real-world applicability, aiming not only to generate new scientific knowledge but also to build early-stage diagnostic strategies and foundational technologies with long-term translational potential.
