Caffeine‐Operated Synthetic Modules for Chemogenetic Control of Protein Activities by Life Style

Abstract

A genetically encoded caffeine‐operated synthetic module (COSMO) is introduced herein as a robust chemically induced dimerization (CID) system. COSMO enables chemogenetic manipulation of biological processes by caffeine and its metabolites, as well as caffeinated beverages, including coffee, tea, soda, and energy drinks. This CID tool, evolved from an anti‐caffeine nanobody via cell‐based high‐throughput screening, permits caffeine‐inducible gating of calcium channels, tumor killing via necroptosis, growth factors‐independent activation of tyrosine receptor kinase signaling, and enhancement of nanobody‐mediated antigen recognition for the severe acute respiratory distress coronavirus 2 (SARS‐CoV‐2) spike protein. Further rationalized engineering of COSMO leads to 34–217‐fold enhancement in caffeine sensitivity (EC50 = 16.9 nanomolar), which makes it among the most potent CID systems like the FK506 binding protein (FKBP)–FKBP rapamycin binding domain (FRB)–rapamycin complex. Furthermore, bivalent COSMO (biCOMSO) connected with a long linker favors intramolecular dimerization and acts as a versatile precision switch when inserted in host proteins to achieve tailored function. Given the modularity and high transferability of COMSO and biCOSMO, these chemical biology tools are anticipated to greatly accelerate the development of therapeutic cells and biologics that can be switched on and off by caffeinated beverages commonly consumed in the daily life.

Application of calcium overload-based ion interference therapy in tumor treatment: strategies, outcomes, and prospects

“…Moreover, considering that the limited penetration of light may restrain its efficacy in deep tumors, some genetic coding tools that utilize chemical factors to manipulate calcium channels have been developed. Wang et al reported a caffeine operated synthesis module (COSMO) that regulates calcium channels through caffeine, its metabolites ( Wang et al, 2021 ). Even more impressively, it also can be activated by caffeinated beverages, including tea, coffee and energy drinks.…”

Programmable synthetic receptors: the next-generation of cell and gene therapies

“… 317 – 319 COSMO-STIM1ct Intracellular soluble ligand Caffeine induces oligomerization of COSMO-STIM1ct, opening ORAI channel to trigger Ca 2+ influx (+) Selectively and remotely control Ca 2+ signal (+) Regulate the function of non-excitable cells (+) Enable orthogonal output function (−) Require addition expression of ORAI Res. 326 Rev. 317 – 319 optoRGK Light Light induces cytosol-to-plasma membrane translocation of engineered RGK GTPases and modulates Ca V channel activity (+) Spatiotemporal control of Ca 2+ signal (−) The efficacy of optoRGK highly depends on Ca V abundancy Res.…”

Rational design of small‐molecule responsive protein switches

“…The binding mode was studied in a crystal structure, revealing a tyrosine‐rich binding pocket and highlighting H‐bonds of Tyr104 of both chains that stabilize the interaction (Lesne et al, 2019 ). A cell‐based high‐throughput screening method was used to further improve the sensitivity of the system, from an EC 50 of 560–100 nM (Wang et al, 2021 ). By now, the caffeine‐controlled nanobody dimerization system has been used in an increasing number of applications spanning from controlling cell receptors (Bojar et al, 2018 ; Chang et al, 2018 ) and dimerization‐dependent enzymes (Scheller et al, 2020 ) to protein re‐localization (Wang et al, 2021 ).…”

“…A cell‐based high‐throughput screening method was used to further improve the sensitivity of the system, from an EC 50 of 560–100 nM (Wang et al, 2021 ). By now, the caffeine‐controlled nanobody dimerization system has been used in an increasing number of applications spanning from controlling cell receptors (Bojar et al, 2018 ; Chang et al, 2018 ) and dimerization‐dependent enzymes (Scheller et al, 2020 ) to protein re‐localization (Wang et al, 2021 ). One concern of this system is that homodimerization systems may be less effective in controlling heterodimerization‐dependent applications, such as two‐hybrid systems for transcription factor‐based inducible gene expression or the assembly of split CARs.…”