Antibody targeting of E3 ubiquitin ligases for receptor degradation

• Published in: Nature (London) 2022-10, Vol.610 (7930), p.182-189
• Main Authors: Marei, Hadir, Tsai, Wen-Ting K., Kee, Yee-Seir, Ruiz, Karen, He, Jieyan, Cox, Chris, Sun, Tao, Penikalapati, Sai, Dwivedi, Pankaj, Choi, Meena, Kan, David, Saenz-Lopez, Pablo, Dorighi, Kristel, Zhang, Pamela, Kschonsak, Yvonne T., Kljavin, Noelyn, Amin, Dhara, Kim, Ingrid, Mancini, Andrew G., Nguyen, Thao, Wang, Chunling, Janezic, Eric, Doan, Alexander, Mai, Elaine, Xi, Hongkang, Gu, Chen, Heinlein, Melanie, Biehs, Brian, Wu, Jia, Lehoux, Isabelle, Harris, Seth, Comps-Agrar, Laetitia, Seshasayee, Dhaya, de Sauvage, Frederic J., Grimmer, Matthew, Li, Jing, Agard, Nicholas J., de Sousa e Melo, Felipe
• Format: Article
• Language: English
• Subjects: 13/106; 13/31; 631/61/51; 631/67/1504/1885/1393; 82/1; 96/95; Antibodies; Binding; Bioavailability; Biodegradation; Cell surface; Colorectal cancer; Colorectal carcinoma; Degradation; Enzymatic activity; Gene expression; Growth factors; Humanities and Social Sciences; Kinases; Ligands; Membrane proteins; multidisciplinary; Permeability; Proteins; Proteolysis; Receptor mechanisms; Receptors; Science; Science (multidisciplinary); Stem cells; Tumors; Ubiquitin; Ubiquitin-protein ligase; Wnt protein
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/s41586-022-05235-6

Most current therapies that target plasma membrane receptors function by antagonizing ligand binding or enzymatic activities. However, typical mammalian proteins comprise multiple domains that execute discrete but coordinated activities. Thus, inhibition of one domain often incompletely suppresses the function of a protein. Indeed, targeted protein degradation technologies, including proteolysis-targeting chimeras 1 (PROTACs), have highlighted clinically important advantages of target degradation over inhibition 2 . However, the generation of heterobifunctional compounds binding to two targets with high affinity is complex, particularly when oral bioavailability is required 3 . Here we describe the development of proteolysis-targeting antibodies (PROTABs) that tether cell-surface E3 ubiquitin ligases to transmembrane proteins, resulting in target degradation both in vitro and in vivo. Focusing on zinc- and ring finger 3 (ZNRF3), a Wnt-responsive ligase, we show that this approach can enable colorectal cancer-specific degradation. Notably, by examining a matrix of additional cell-surface E3 ubiquitin ligases and transmembrane receptors, we demonstrate that this technology is amendable for ‘on-demand’ degradation. Furthermore, we offer insights on the ground rules governing target degradation by engineering optimized antibody formats. In summary, this work describes a strategy for the rapid development of potent, bioavailable and tissue-selective degraders of cell-surface proteins. Membrane-bound E3 ubiquitin ligases RNF43 and ZNRF3 are overexpressed in colorectal cancer, and can be repurposed using proteolysis-targeting antibodies (PROTABs) to selectively degrade cell-surface receptors in tumours.