When Light Meets Chemok ine Receptors: Photocontrolling ACKR3 and CXCR4 Function
Photo pharmacology integrates photochemical principles with pharmacology to achieve light-dependent regulation of biological targets using photo switchable molecules. Light-driven control enables temporal and spati al accuracy, offering new possibilities to study dynamic signaling pr ocesses while limiting undesired systemic effects.G protein-coupled r eceptors (GPCRs) represent the largest group of proteins targeted by therapeutic drugs and play central roles in numerous physiological fu nctions. Within GPCRs, the chemokine receptors CXCR4 and ACKR3 are cr itically involved in immune cell trafficking, tissue development, inf lammation, and cancer progression. However, conventional pharmacologi cal tools targeting these receptors act globally and lack the ability to reversibly modulate receptor activity in space and time. This PhD thesis reports the development and pharmacological characterization of azobenzene-based photoswitchable ligands designed to enable optica l control of CXCR4 and ACKR3. To support ligand discovery and evaluat ion, chemokine receptor binding assays based on NanoBRET technology w ere established and expanded into a multiplexed format. Using these p latforms, the first photoswitchable CXCR4 antagonist was identified, alongside the first photoswitchable agonist and inverse agonist for A CKR3, allowing reversible and bidirectional modulation of receptor ac tivity with light. The compounds were characterized using multiple GP CR-relevant assays, including binding studies, β-arrestin2 recruitme nt assays, FRET-based biosensors, and fluorescence imaging of ligand binding. In addition, assay formats suitable for in situ photoswitchi ng were assessed and adapted. Overall, this work demonstrates that ch emokine receptor activity can be modulated with high spatiotemporal p recision using photoswitchable ligands. The tools and methodologies d eveloped in this thesis expand the photopharmacological toolkit for G PCR research and provide new opportunities to investigate CXCR4 and A CKR3 signaling dynamics, with potential relevance for future light-co ntrolled therapeutic strategies.
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DESCRIPTION: Photopharmacology integrates photochemical principles wit h pharmacology to achieve light-dependent regulation of biological ta rgets using photoswitchable molecules. Light-driven control enables t emporal and spatial accuracy, offering new possibilities to study dyn amic signaling processes while limiting undesired systemic effects.G protein-coupled receptors (GPCRs) represent the largest group of prot eins targeted by therapeutic drugs and play central roles in numerous physiological functions. Within GPCRs, the chemokine receptors CXCR4 and ACKR3 are critically involved in immune cell trafficking, tissue development, inflammation, and cancer progression. However, conventi onal pharmacological tools targeting these receptors act globally and lack the ability to reversibly modulate receptor activity in space a nd time. This PhD thesis reports the development and pharmacological characterization of azobenzene-based photoswitchable ligands designed to enable optical control of CXCR4 and ACKR3. To support ligand disc overy and evaluation, chemokine receptor binding assays based on Nano BRET technology were established and expanded into a multiplexed form at. Using these platforms, the first photoswitchable CXCR4 antagonist was identified, alongside the first photoswitchable agonist and inve rse agonist for ACKR3, allowing reversible and bidirectional modulati on of receptor activity with light. The compounds were characterized using multiple GPCR-relevant assays, including binding studies, β-ar restin2 recruitment assays, FRET-based biosensors, and fluorescence i maging of ligand binding. In addition, assay formats suitable for in situ photoswitching were assessed and adapted. Overall, this work dem onstrates that chemokine receptor activity can be modulated with high spatiotemporal precision using photoswitchable ligands. The tools an d methodologies developed in this thesis expand the photopharmacologi cal toolkit for GPCR research and provide new opportunities to invest igate CXCR4 and ACKR3 signaling dynamics, with potential relevance fo r future light-controlled therapeutic strategies. More information on the thesis When Light Meets Chemokine Receptors: Photocontro lling ACKR3 and CXCR4 Function END:VEVENT END:VCALENDAR