Programming T cells for Early Cancer Detection with Customized Protease-Activatable Receptors

Phuengkham, H.; Chen, Y.; Sivakumar, A.; Zamat, A. H.; Gamboa, L.; Mac, Q. D.; Lee, H. J.; Rogers, L. C.; You, J.; Steele, S. A.; Zhu, S.; Gollins, M. S.; Blazeck, J.; Qiu, P.; Kwong, G. A.

doi:10.64898/2026.01.16.699939

bioRxiv · 10.64898/2026.01.16.699939

Programming T cells for Early Cancer Detection with Customized Protease-Activatable Receptors

Phuengkham, H.·Chen, Y.·Sivakumar, A.·Zamat, A. H.·Gamboa, L.·Mac, Q. D.·Lee, H. J.·Rogers, L. C.·You, J.·Steele, S. A.·Zhu, S.·Gollins, M. S.·Blazeck, J.·Qiu, P.·Kwong, G. A.

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Abstract

Early cancer detection has the potential to reduce cancer mortality, yet endogenous tumor-shed biomarkers lack sensitivity for early-stage disease. We report OncoSCOUT, a cancer detection strategy using T cells engineered with protease-activatable receptors (PARs) that conditionally recognize tumor cells and release a synthetic biomarker for detection in urine. These PARs comprise masked synthetic Notch receptors in which antigen binding is blocked by a peptide mimotope tethered via a protease-cleavable linker. We demonstrate that requiring both extracellular protease activity and tumor antigen recognition improves spatial specificity and minimizes off-tumor activation of PAR T cells in vivo. To identify tumor-selective PARs, we adoptively transferred a HER2-targeted PAR library displaying [~]160,000 unique 4-mer amino acid linkers and discovered multiple variants significantly enriched in a HER2-positive cancer xenograft model. Using a single customized PAR, we show that OncoSCOUT can detect total tumor burdens as small as 10-30 mm3 with significantly improved sensitivity than the protein biomarker CA 15-3 or a 20-plex circulating tumor DNA (ctDNA) assay.

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Phuengkham, H., Chen, Y., Sivakumar, A., Zamat, A. H., Gamboa, L., Mac, Q. D., Lee, H. J., Rogers, L. C., You, J., Steele, S. A., Zhu, S., Gollins, M. S., Blazeck, J., Qiu, P., Kwong, G. A.. 2026-01-20. Programming T cells for Early Cancer Detection with Customized Protease-Activatable Receptors. https://doi.org/10.64898/2026.01.16.699939

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