Nanomedicines targeting protease-activated receptor 2 in endosomes provide sustained analgesia

作者：Shavonne Teng, Rocco Latorre, Divya Bhansali, Parker K. Lewis, Rachel E. Pollard, Chloe J. Peach, Badr Sokrat, Gokul Sriman Thanigai Arasu, Tracy Chiu, Paz Duran, Nestor N. Jiménez-Vargas, Abby Mocherniak, Matthew Bogyo, Michael Gaspari, Stephen Vanner, Nathalie M. Pinkerton, Kam W. Leong, Brian L. Schmidt, Dane D. Jensen, Nigel W. Bunnett · 发表于：Proceedings of the National Academy of Sciences · 年份：2025 · DOI：10.1073/pnas.2412687122 · 被引用次数：8 · 研究领域：Lipid Membrane Structure and Behavior、Receptor Mechanisms and Signaling、Neuropeptides and Animal Physiology

Although many internalized G protein-coupled receptors (GPCRs) continue to signal, the mechanisms and outcomes of intracellular GPCR signaling are uncertain due to the challenges of measuring organelle-specific signals and of selectively antagonizing receptors in intracellular compartments. Herein, genetically encoded biosensors targeted to the plasma membrane and early endosomes were used to analyze compartmentalized signaling of protease-activated receptor 2 (PAR 2 ); the propensity of nanoparticles (NPs) to accumulate in endosomes was leveraged to preferentially antagonize intracellular PAR 2 signaling of pain. PAR 2 agonists evoked sustained activation of PAR 2 , Gαq, and β-arrestin-1 in early endosomes and activated extracellular signal regulated kinase (ERK) in the cytosol and nucleus, measured with targeted biosensors. Fluorescent dendrimer and core-shell polymeric NPs accumulated in endosomes of HEK293T cells, colonic epithelial cells, and nociceptors, detected by confocal microscopy. NPs efficiently encapsulated and slowly released AZ3451, a negative allosteric PAR 2 modulator. NP-encapsulated AZ3451, but not unencapsulated AZ3451, rapidly and completely reversed PAR 2 , Gαq, and β-arrestin-1 activation in early endosomes and ERK activation in the cytosol and nucleus. When administered into the mouse colon lumen, fluorescent dendrimer NPs accumulated in endosomes of colonocytes and polymeric NPs accumulated in neurons, sites of PAR 2 expression. Both NP formulations ...