Small molecule-nanobody conjugate induced proximity controls intracellular processes and modulates endogenous unligandable targets.
Xiaofeng SunChengjian ZhouSimin XiaXi ChenPublished in: Nature communications (2023)
Chemically induced proximity (CIP) is a powerful tool to study cellular functions. However with current CIP inducers it is difficult to directly modulate unligandable and endogenous targets, and therapeutic translational potential is also restricted. Herein, we combine CIP and chemical nanobody engineering and create cell-permeable small molecule-nanobody conjugate inducers of proximity (SNACIPs). The SNACIP inducer cRGT carrying a cyclic cell-penetrating peptide rapidly enters live cells and dimerizes eDHFR and GFP-variants. cRGT enables minute-scale, reversible, no-wash and dose-dependent control of cellular processes including signaling cascade, cargo transport and ferroptosis. Small-molecule motifs can also be installed via post-translational modifications. Therefore, latent-type SNACIPs including cRTC are designed that are functionally assembled inside living cells. cRTC contains a nanobody against an intrinsically disordered protein TPX2, a microtubule nucleation factor overexpressed in various cancers. Cancer cell proliferation is inhibited and tumor growth is suppressed in vivo. Hence, SNACIPs are valuable proximity inducers for regulating cellular functions.
Keyphrases
- small molecule
- protein protein
- living cells
- cell proliferation
- high glucose
- single cell
- diabetic rats
- fluorescent probe
- cell therapy
- induced apoptosis
- cell death
- single molecule
- drug induced
- stem cells
- cell cycle arrest
- oxidative stress
- copy number
- cell cycle
- gene expression
- mesenchymal stem cells
- endothelial cells
- genome wide
- childhood cancer
- amino acid
- dna methylation
- reactive oxygen species