Conditionally Controlling Human TLR2 Activity via Trans-Cyclooctene Caged Ligands

Bioconjugate Chemistry, vol. 31, pp. 1685–1692

Abstract

Toll-like receptors (TLRs) are key pathogen sensors of the immune system. Their activation results in the production of cytokines, chemokines, and costimulatory molecules that are crucial for innate and adaptive immune responses. In recent years, specific (sub)-cellular location and timing of TLR activation have emerged as parameters for defining the signaling outcome and magnitude. To study the subtlety of this signaling, we here report a new molecular tool to control the activation of TLR2 via "click-to-release"-chemistry. We conjugated a bioorthogonal trans-cyclooctene (TCO) protecting group via solid support to a critical position within a synthetic TLR2/6 ligand to render the compound unable to initiate signaling. The TCO-group could then be conditionally removed upon addition of a tetrazine, resulting in restored agonist activity and TLR2 activation. This approach was validated on RAW264.7 macrophages and various murine primary immune cells as well as human cell line systems, demonstrating that TCO-caging constitutes a versatile approach for generating chemically controllable TLR2 agonists.

Authors 8

1. Michel J. van de Graaff Aachen Department of Bio-organic Synthesis

   Leiden University

   Affiliation as printed

   Leiden University , , Einsteinweg 55 , , ,

   Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, Zuid-Holland, The Netherlands

2. Timo Oosenbrug Aachen Department of Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center , , Einthovenweg 20 , , ,

   Department of Cell and Chemical Biology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, Zuid-Holland, The Netherlands

3. Mikkel H. S. Marqvorsen Aachen Department of Bio-organic Synthesis

   Leiden University

   Affiliation as printed

   Leiden University , , Einsteinweg 55 , , ,

   Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, Zuid-Holland, The Netherlands

4. Clarissa Rodrigues Nascimento

   Hôpital Necker-Enfants Malades · Centre National de la Recherche Scientifique · Inserm · Université Paris Cité · Institut Necker Enfants Malades

   Affiliation as printed

   Université de Paris , , Faculté de Médecine, 156 Rue de Vaugirard , ,

   INEM, INSERM, Unité 1151-CNRS UMR 8253, Université de Paris, Faculté de Médecine, 156 Rue de Vaugirard, 75015 Paris, France

   Institut Necker Enfants-Malades

5. Mark A. R. de Geus Aachen Department of Bio-organic Synthesis

   Leiden University

   Affiliation as printed

   Leiden University , , Einsteinweg 55 , , ,

   Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, Zuid-Holland, The Netherlands

6. Bénédicte Manoury

   Hôpital Necker-Enfants Malades · Centre National de la Recherche Scientifique · Inserm · Université Paris Cité · Institut Necker Enfants Malades

   Affiliation as printed

   Université de Paris , , Faculté de Médecine, 156 Rue de Vaugirard , ,

   INEM, INSERM, Unité 1151-CNRS UMR 8253, Université de Paris, Faculté de Médecine, 156 Rue de Vaugirard, 75015 Paris, France

   Institut Necker Enfants-Malades

7. Maaike E. Ressing corresponding Aachen Department of Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center , , Einthovenweg 20 , , ,

   Department of Cell and Chemical Biology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, Zuid-Holland, The Netherlands

8. Sander I. van Kasteren corresponding Aachen Department of Bio-organic Synthesis

   Leiden University

   Affiliation as printed

   Leiden University , , Einsteinweg 55 , , ,

   Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, Zuid-Holland, The Netherlands