RT Journal Article SR Electronic T1 Harnessing PROTAC technology to combat stress hormone receptor activation JF bioRxiv FD Cold Spring Harbor Laboratory SP 2023.03.17.533120 DO 10.1101/2023.03.17.533120 A1 Mahshid Gazorpak A1 Karina M. Hugentobler A1 Dominique Paul A1 Pierre-Luc Germain A1 Kei Matthis A1 Remo Rudolf A1 Sergio Mompart Barrenechea A1 Miriam Kretschmer A1 Vincent Fischer A1 Xiaohan Xue A1 Mattia Privitera A1 Iryna Ivanova A1 Andreas Hierlemann A1 Onno C. Meijer A1 Erick M. Carreira A1 Johannes Bohacek A1 Katharina Gapp YR 2023 UL http://biorxiv.org/content/early/2023/03/18/2023.03.17.533120.abstract AB Counteracting the overactivation of glucocorticoid receptors (GR) is an important therapeutic goal in stress-related psychiatry and beyond. The only clinically approved GR antagonist lacks selectivity and induces unwanted side effects. To complement existing tools of small-molecule-based inhibitors, we present a highly potent, novel catalytically-driven GR degrader, KH-103, based on proteolysis-targeting chimera technology. This selective degrader enables immediate and reversible GR depletion that is independent of genetic manipulation and circumvents transcriptional adaptations to inhibition. KH-103 achieves passive inhibition, preventing agonistic induction of gene expression, and significantly averts the GR’s genomic effects compared to two currently available inhibitors. Application in primary-neuron cultures revealed the dependency of a glucocorticoid-induced increase in spontaneous calcium activity on GR. Finally, we present a proof of concept for application in-vivo. KH-103 opens opportunities for a more lucid interpretation of GR functions with translational potential.Competing Interest StatementThe authors have declared no competing interest.AAVAdeno-associated virusALSAmyotrophic Lateral SclerosisBBBBlood-brain-barrierBpBase pairChIP-seqChromatin immunoprecipitation sequencingCPMCounts per millionCRBNCereblonCypsCytochromes P450DEGDifferentially expressed geneDEXDexamethasoneDMSODimethyl sulfoxidedTAGDegradation TAGDusp1Dual-specificity phosphatase 1EGFPEnhanced green fluorescent proteinFDRFalse discovery rateFKBP12FK506-binding protein 12FKBP5FK506-binding Protein 51GCGlucocorticoidGRGlucocorticoid receptorHHour(s)HEK293Human embryonic kidney 293Histone 3H3HPAHypothalamus-Pituitary-AdrenalHzHertzIL-6Interleukin-6KDDissociation constantKdaKilodaltonLBDLigand binding domainlogFCLog fold changeMAP2Microtubule-associated protein 2MIFMifepristoneMinMinute(s)MProSARS-CoV-2 main proteaseMRMineralocorticoid receptorN2aNeuro 2aPDBProtein data bankPEGPoly(ethylene glycol)Per1Period circadian regulator 1PROTACProteolysis targeting chimerasPXRPregnane X receptorRT-qPCRReverse transcription-quantitative polymerase chain reactionSgk1Serum/glucocorticoid regulated kinase 1TAUTubulin-associated unitTSSTranscription start siteUPSUbiquitin-proteasome systemVDACVoltage-dependent anion channelAAVAdeno-associated virusALSAmyotrophic Lateral SclerosisBBBBlood-brain-barrierBpBase pairChIP-seqChromatin immunoprecipitation sequencingCPMCounts per millionCRBNCereblonCypsCytochromes P450DEGDifferentially expressed geneDEXDexamethasoneDMSODimethyl sulfoxidedTAGDegradation TAGDusp1Dual-specificity phosphatase 1EGFPEnhanced green fluorescent proteinFDRFalse discovery rateFKBP12FK506-binding protein 12FKBP5FK506-binding Protein 51GCGlucocorticoidGRGlucocorticoid receptorHHour(s)HEK293Human embryonic kidney 293Histone 3H3HPAHypothalamus-Pituitary-AdrenalHzHertzIL-6Interleukin-6KDDissociation constantKdaKilodaltonLBDLigand binding domainlogFCLog fold changeMAP2Microtubule-associated protein 2MIFMifepristoneMinMinute(s)MProSARS-CoV-2 main proteaseMRMineralocorticoid receptorN2aNeuro 2aPDBProtein data bankPEGPoly(ethylene glycol)Per1Period circadian regulator 1PROTACProteolysis targeting chimerasPXRPregnane X receptorRT-qPCRReverse transcription-quantitative polymerase chain reactionSgk1Serum/glucocorticoid regulated kinase 1TAUTubulin-associated unitTSSTranscription start siteUPSUbiquitin-proteasome systemVDACVoltage-dependent anion channel