PT  - JOURNAL ARTICLE
AU  - Kayvon Pedram
AU  - D. Judy Shon
AU  - Gabrielle S. Tender
AU  - Natalia R. Mantuano
AU  - Jason J. Northey
AU  - Kevin J. Metcalf
AU  - Simon P. Wisnovsky
AU  - Nicholas M. Riley
AU  - Giovanni C. Forcina
AU  - Stacy A. Malaker
AU  - Angel Kuo
AU  - Benson M. George
AU  - Caitlyn L. Miller
AU  - Kerriann M. Casey
AU  - José G. Vilches-Moure
AU  - Valerie M. Weaver
AU  - Heinz Laübli
AU  - Carolyn R. Bertozzi
TI  - Design of a mucin-selective protease for targeted degradation of cancer-associated mucins
AID  - 10.1101/2022.05.20.492748
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.05.20.492748
4099  - http://biorxiv.org/content/early/2022/05/20/2022.05.20.492748.short
4100  - http://biorxiv.org/content/early/2022/05/20/2022.05.20.492748.full
AB  - Targeted protein degradation is an emerging strategy for the elimination of classically undruggable proteins. Here, to expand the landscape of substrates that can be selectively degraded, we designed degraders which are dependent on both peptide sequence and glycosylation status of the target protein. We applied this approach to mucins, O-glycosylated proteins that drive cancer progression through biophysical and immunological mechanisms. Engineering of a bacterial mucin-selective protease yielded a variant for fusion to a cancer antigen-binding nanobody. The resulting conjugate selectively degraded mucins on cancer cells, promoted cell death in culture models of mucin-driven growth and survival, and reduced tumor growth in murine models of breast cancer progression. This work establishes a blueprint for the development of biologics which degrade specific glycoforms of cell surface proteins.Competing Interest StatementA patent application relating to the use of targeted enzymes to digest mucin-domain glycoproteins has been filed by Stanford University (docket no. STAN-1929PRV). C.R.B. is a co-founder and scientific advisory board member of Lycia Therapeutics, Palleon Pharmaceuticals, Enable Bioscience, Redwood Biosciences (a subsidiary of Catalent), OliLux Bio, Grace Science LLC, and InterVenn Biosciences. H.L. received travel grants and consultant fees from Bristol-Myers Squibb (BMS) and Merck, Sharp and Dohme (MSD), Roche, InterVenn, and Alector. H.L. received research support from BMS, Novartis, GlycoEra, and Palleon Pharmaceuticals.