When Light Meets Che mokine Receptors: Photocontrolling ACKR3 and CXCR4 Function
< p>Photopharmacology integrates photochemical principles with pharmaco logy to achieve light-dependent regulation of biological targets usin g photoswitchable molecules. Light-driven control enables temporal an d spatial accuracy, offering new possibilities to study dynamic signa ling processes while limiting undesired systemic effects.G protein-co upled receptors (GPCRs) represent the largest group of proteins targe ted by therapeutic drugs and play central roles in numerous physiolog ical functions. Within GPCRs, the chemokine receptors CXCR4 and ACKR3 are critically involved in immune cell trafficking, tissue developme nt, inflammation, and cancer progression. However, conventional pharm acological tools targeting these receptors act globally and lack the ability to reversibly modulate receptor activity in space and time. T his PhD thesis reports the development and pharmacological characteri zation of azobenzene-based photoswitchable ligands designed to enable optical control of CXCR4 and ACKR3. To support ligand discovery and evaluation, chemokine receptor binding assays based on NanoBRET techn ology were established and expanded into a multiplexed format. Using these platforms, the first photoswitchable CXCR4 antagonist was ident ified, alongside the first photoswitchable agonist and inverse agonis t for ACKR3, allowing reversible and bidirectional modulation of rece ptor activity with light. The compounds were characterized using mult iple GPCR-relevant assays, including binding studies, β-arrestin2 re cruitment assays, FRET-based biosensors, and fluorescence imaging of ligand binding. In addition, assay formats suitable for in situ photo switching were assessed and adapted. Overall, this work demonstrates that chemokine receptor activity can be modulated with high spatiotem poral precision using photoswitchable ligands. The tools and methodol ogies developed in this thesis expand the photopharmacological toolki t for GPCR research and provide new opportunities to investigate CXCR 4 and ACKR3 signaling dynamics, with potential relevance for future l ight-controlled therapeutic strategies.More information on the thesis
DESCRIPTION: When Light Meets Chemokine Receptors: Photocontrolling AC KR3 and CXCR4 Function Photopharmacology integrates photochemical pri nciples with pharmacology to achieve light-dependent regulation of bi ological targets using photoswitchable molecules. Light-driven contro l enables temporal and spatial accuracy, offering new possibilities t o study dynamic signaling processes while limiting undesired systemic effects.G protein-coupled receptors (GPCRs) represent the largest gr oup of proteins targeted by therapeutic drugs and play central roles in numerous physiological functions. Within GPCRs, the chemokine rece ptors CXCR4 and ACKR3 are critically involved in immune cell traffick ing, tissue development, inflammation, and cancer progression. Howeve r, conventional pharmacological tools targeting these receptors act g lobally and lack the ability to reversibly modulate receptor activity in space and time. This PhD thesis reports the development and pharm acological characterization of azobenzene-based photoswitchable ligan ds designed to enable optical control of CXCR4 and ACKR3. To support ligand discovery and evaluation, chemokine receptor binding assays ba sed on NanoBRET technology were established and expanded into a multi plexed format. Using these platforms, the first photoswitchable CXCR4 antagonist was identified, alongside the first photoswitchable agoni st and inverse agonist for ACKR3, allowing reversible and bidirection al modulation of receptor activity with light. The compounds were cha racterized using multiple GPCR-relevant assays, including binding stu dies, β-arrestin2 recruitment assays, FRET-based biosensors, and flu orescence imaging of ligand binding. In addition, assay formats suita ble for in situ photoswitching were assessed and adapted. Overall, th is work demonstrates that chemokine receptor activity can be modulate d with high spatiotemporal precision using photoswitchable ligands. T he tools and methodologies developed in this thesis expand the photop harmacological toolkit for GPCR research and provide new opportunitie s to investigate CXCR4 and ACKR3 signaling dynamics, with potential r elevance for future light-controlled therapeutic strategies.More info rmation on the thesis END:VEVENT END:VCALENDAR