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The Spatiotemporal Bias in Proinflammatory p38 Signaling Revealed by a Forster's Resonance Energy Transfer (FRET)‐based Platform for Mapping GPCR‐induced Vascular Inflammation
Author(s) -
Burton Jeremy,
Okalova Jennifer,
Grimsey Neil
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.02580
Subject(s) - microbiology and biotechnology , g protein coupled receptor , p38 mitogen activated protein kinases , signal transduction , inflammation , proinflammatory cytokine , biology , mapk/erk pathway , immunology
Mitogen‐activated protein kinase (MAPK) p38 is a critical pro‐inflammatory signaling protein associated with pulmonary vascular disease such as acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disorder (COPD), Sars‐Cov‐2, and is a key therapeutic target. However, p38‐targeted therapeutics have largely failed in clinical trials in part due to the ubiquitous expression and essential physiological roles of p38 activity in almost all tissues. We have previously shown that G‐protein coupled receptor (GPCR) stimulation activates an understudied atypical p38 pathway, through direct interaction between p38 and an essential adaptor protein TGF‐β Activated Kinase 1 Binding Protein 1 (TAB1), inducing p38 autophosphorylation. There is a critical gap in our understanding of how this pathway induces inflammation, and a greater understanding of atypical p38 signaling could provide alternative therapeutic approaches. We hypothesized that the spatiotemporal regulation of atypical p38 signaling is critical for downstream signaling responses associated with pulmonary inflammation. In this work, we have developed a Forster's Resonance Energy Transfer (FRET) platform of sub‐cellular localized biosensors to map p38 activity. Through exploration of GPCRs known to activate atypical p38 signaling, we demonstrate that GPCRs significantly alter the dynamics of p38 activity in part through sequestration of the kinase in the cytosol. This is contrary to rapid nuclear translocation after induction of classical MKK3/6 mediated p38 signaling. This ligand‐dependent bias in the spatiotemporal dynamics of p38 activity represents an under‐explored mechanism for regulation of vascular inflammation. Our current studies are focused on defining how cytosolic atypical p38 modulates pulmonary inflammatory signaling. We have established a basis for further expansion of this system to differentiate cytosolic‐biased GPCR signaling at the endosome and will adapt this platform of biosensors for further exploration in pulmonary vascular cells. Combined, these studies provide foundational insight into the spatiotemporal bias of TAB1‐mediated p38 signaling and its modulation of vascular inflammatory disease.