Engineering Biomimetic Trogocytosis with Farnesylated Chemically Self-Assembled Nanorings

Abstract

Inspired by the natural intercellular material-transfer process of trans-endocytosis or trogocytosis, we proposed that targeted farnesylated chemically self-assembled nanorings (f-CSANs) could serve as a biomimetic trogocytosis vehicle for engineering directional cargo transfer between cells, thus allowing cell–cell interactions to be monitored and facilitating cell–cell communications. The membranes of sender cells were stably modified by hydrophobic insertion with the targeted f-CSANs, which were efficiently transferred to receiver cells expressing the appropriate receptors by endocytosis. CSAN-assisted cell–cell cargo transfer (C4T) was demonstrated to be receptor specific and dependent on direct cell–cell interactions, the rate of receptor internalization, and the level of receptor expression. In addition, C4T was shown to facilitate cell-to-cell delivery of an apoptosis inducing drug, as wells as antisense oligonucleotides. Taken together, the C4T approach is a potentially versatile biomimetic trogocytosis platform that can be deployed as a macro-chemical biological tool for monitoring cell–cell interactions and engineering cell–cell communications.

Targeted Drug Delivery by MMAE Farnesyl-Bioconjugated Multivalent Chemically Self-Assembled Nanorings Induces Potent Receptor-Dependent Immunogenic Cell Death

“…The E1-DHFR 2 -MMAE CSANs were shown to be efficiently internalized by A431-R cells through specific EGFR binding, which was confirmed by the significantly higher fluorescent CSAN signal when the cells were treated at 37 °C compared with 4 °C. A significant reduction in fluorescent CSAN internalization was observed when the cells were incubated in the presence of unlabeled anti-EGFR CSANs, consistent with previous studies of anti-EGFR CSAN target specificity (Figure b). , Additionally, the surface binding and internalization of the E1-DHFR 2 -MMAE CSANs were also imaged by fluorescence microscopy. Those studies revealed that the CSANs underwent cellular uptake by the EGFR + A431-R cells, indicated by the green punctate spots from the CSANs within the cells.…”

“…An anti-EGFR Fn3 domain generated through yeast surface display and screening was genetically fused to the N-terminus of the DHFR 2 fusion protein to serve as the targeting ligand. ,, A C-terminal CVIA sequence incorporated into the DHFR 2 fusion proteins was also previously demonstrated to be efficiently recognized and prenylated with various farnesyl diphosphate derivatives. − To afford bioconjugation, the E1-DHFR 2 -CVIA monomer was efficiently farnesylated with an azide-containing farnesyl analog (C10-N 3 -OPP) by farnesyltransferase . The farnesylated E1-DHFR 2 -CVIA protein (E1-DHFR 2 -N 3 ) was subsequently conjugated with the DBCO-functionalized prodrug DBCO-PEG 4 -VC-PAB-MMAE (DBCO-MMAE) through a strain-promoted click reaction to prepare the desired E1-DHFR 2 -MMAE protein-drug conjugate .…”

“…22−24 To afford bioconjugation, the E1-DHFR 2 -CVIA monomer was efficiently farnesylated with an azide-containing farnesyl analog (C10-N 3 -OPP) by farnesyltransferase. 25 The farnesylated E1-DHFR 2 -CVIA protein (E1-DHFR 2 -N 3 ) was subsequently conjugated with the DBCO-functionalized prodrug DBCO-PEG 4 -VC-PAB-MMAE (DBCO-MMAE) through a strain-promoted click reaction to prepare the desired E1-DHFR 2 -MMAE protein-drug conjugate. 26 Formation of E1-DHFR 2 -MMAE was confirmed by LC-MS (Figure 2a).…”

Multivalent Fluorinated Nanorings for On-Cell 19F NMR

“…Given the modularity of this design, fluorinated imaging tags could also be applied to targeting a variety of other extracellular receptors, including those with significantly slower rates of internalization. EpCAM is one such alternative, which we have shown to internalize 24-fold slower than EGFR when targeting with CSANs while maintaining high expression on a variety of solid tumor cells …”

“…EpCAM is one such alternative, which we have shown to internalize 24-fold slower than EGFR when targeting with CSANs while maintaining high expression on a variety of solid tumor cells. 38 ■ ASSOCIATED CONTENT * sı Supporting Information…”