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Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

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Engineering reversible cell-cell interactions using enzymatically lipidated chemically self-assembled nanorings.

Yiao Wang¹, Ozgun Kilic², Clifford M Csizmar²

¹Department of Chemistry, University of Minnesota Minneapolis Minnesota 55455 USA diste001@umn.edu wagne003@umn.edu.

Chemical Science

|June 25, 2021

Summary

This summary is machine-generated.

Researchers developed chemically self-assembled nanorings (CSANs) to control cell-cell interactions for therapies. These nanorings target specific cells and can be rapidly disassembled, enabling precise control over cell interactions and T cell-mediated cytotoxicity.

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Area of Science:

Biotechnology and Bioengineering
Cellular and Molecular Biology
Immunology

Background:

Multicellular biology relies on controlled cell-cell interactions.
Engineering cell-based therapies requires precise manipulation of these interactions.
Existing methods may lack temporal control or specificity.

Purpose of the Study:

To develop a novel multivalent, lipidated scaffold for rapid and reversible manipulation of cell-cell interactions.
To engineer antigen-targeted nanostructures for specific cell engagement and therapeutic applications.
To investigate the potential of these nanostructures in T cell-mediated cytotoxicity.

Main Methods:

Formation of chemically self-assembled nanorings (CSANs) via bivalent dihydrofolate reductase (DHFR^2) fusion proteins and a chemical dimerizer.
Incorporation of targeting proteins (anti-EGFR, anti-EpCAM) and a CAAX-box sequence into DHFR^2 monomers.
Enzymatic prenylation of monomers followed by assembly into CSANs, with subsequent cell surface modification and characterization.
Assessment of CSAN disassembly using trimethoprim and evaluation of T cell-mediated cytotoxicity against target cells.

Main Results:

Prenylated CSANs efficiently modified lymphocyte cell surfaces, exhibiting a half-life of over 3 days and associating with lipid rafts.
Antigen-targeted CSANs specifically interacted with target cells, and trimethoprim treatment rapidly disassembled the nanostructures, terminating interactions.
Activated PBMCs modified with prenylated CSANs induced selective cytotoxicity toward EGFR-expressing cells within 2 hours, independent of CD3 engagement.

Conclusions:

Antigen-targeted prenylated CSANs offer a versatile platform for regulating specific cell-cell interactions.
The rapid and reversible nature of CSANs provides temporal control crucial for studying cell interactions and T cell-mediated cytotoxicity.
This approach holds significant potential for fundamental research and therapeutic applications in cell-based therapies.