6XSW

Structure of the Notch3 NRR in complex with an antibody Fab Fragment

Summary for 6XSW

• Entry DOI: 10.2210/pdb6xsw/pdb
• Descriptor: Anti-N3 Fab Heavy Chain, Anti-N3 Fab Light Chain, Neurogenic locus notch homolog protein 3, ... (7 entities in total)
• Functional Keywords: notch3, antibody, signaling protein, signaling protein-immune system complex, signaling protein/immune system
• Biological source: Rattus norvegicus; More
• Total number of polymer chains: 12
• Total formula weight: 318978.95

Authors

Bard, J. (deposition date: 2020-07-16, release date: 2021-07-21, Last modification date: 2024-11-06)

Primary citation

Geles, K.G.,Gao, Y.,Giannakou, A.,Sridharan, L.,Yamin, T.T.,Zhang, J.,Karim, R.,Bard, J.,Piche-Nicholas, N.,Charati, M.,Maderna, A.,Lucas, J.,Golas, J.,Guffroy, M.,Pirie-Shepherd, S.,Roy, M.,Qian, J.,Franks, T.,Zhong, W.,O'Donnell, C.J.,Tchistiakova, L.,Gerber, H.P.,Sapra, P.
NOTCH3-targeted antibody drug conjugates regress tumors by inducing apoptosis in receptor cells and through transendocytosis into ligand cells.
Cell Rep Med, 2:100279-100279, 2021

PubMed Abstract: Aberrant NOTCH3 signaling and overexpression is oncogenic, associated with cancer stem cells and drug resistance, yet therapeutic targeting remains elusive. Here, we develop NOTCH3-targeted antibody drug conjugates (NOTCH3-ADCs) by bioconjugation of an auristatin microtubule inhibitor through a protease cleavable linker to two antibodies with differential abilities to inhibit signaling. The signaling inhibitory antibody rapidly induces ligand-independent receptor clustering and internalization through both caveolin and clathrin-mediated pathways. The non-inhibitory antibody also efficiently endocytoses via clathrin without inducing receptor clustering but with slower lysosomal co-localization kinetics. In addition, DLL4 ligand binding to the NOTCH3 receptor mediates transendocytosis of NOTCH3-ADCs into ligand-expressing cells. NOTCH3-ADCs internalize into receptor and ligand cells independent of signaling and induce cell death in both cell types representing an atypical mechanism of ADC cytotoxicity. Treatment of xenografts with NOTCH3-ADCs leads to sustained tumor regressions, outperforms standard-of-care chemotherapy, and allows targeting of tumors that overexpress NOTCH3 independent of signaling inhibition.

PubMed: 34095881
DOI: 10.1016/j.xcrm.2021.100279

Experimental method

X-RAY DIFFRACTION (2.98 Å)