Biology subjects

Bronneberg, G.

Publications and source records attributed to Bronneberg, G..

2 recordsLinked to original sources

bioRxiv2025

SMAD2 sets divergent thresholds for TGF-β-induced SMAD1/5 signaling and IgE-mediated pro-inflammatory activation of mast cells

ObjectiveTGF-{beta}/SMAD signaling controls mast cell (MC) development and exerts anti-inflammatory functions, while antigen (Ag)-triggered Fc{varepsilon}RI/MAPK activation commands pro-inflammatory processes. SMAD2 is a signaling hub integrating both, TGF-{beta}/ALK5-mediated SMAD2 C-terminal-and ERK-mediated SMAD2 linker-phosphorylation. Here, we analyzed the role of SMAD2 in TGF-{beta}-and Ag-stimulated signaling, and their cross-talk which regulate MC responses. MethodsThe CRISPR-Cas method was used to disable SMAD2 expression in the PMC-306 cell line. The transcriptome of the corresponding cells was analyzed by NGS in homeostatic and stimulated conditions. Activation of signaling intermediates, target gene responses and effector function were analyzed by qPCR, Western blot and ELISAs. The causal association of SMAD2 with the found effects was proven by re-introduction of a tagged SMAD2 in knock out cells. ResultsThe absence of SMAD2 led to increased proliferation and survival, and decreased transcription of target genes like Smad7 and Jun in steady state and upon TGF-{beta} treatment. Intriguingly, SMAD2 was found to regulate TGF-{beta}-mediated SMAD1/5 activation, resulting in augmented expression of Id3 in SMAD2-deficient MCs. Unexpectedly, we found that SMAD2 is indispensable for Ag-triggered production of pro-inflammatory cytokines, such as IL-6 and TNF. Re-introducing SMAD2 restores these events with varying sensitivity depending on the receptor system triggered. ConclusionsOur findings highlight a previously unrecognized role of SMAD2 as a decisive hub in TGF-{beta}-ALK5-SMAD1/5 and Ag-Fc{varepsilon}RI-MAPK signaling, suggesting that pharmacological modulation of SMAD2 activity could be leveraged as regulator of both suppressive and inflammatory MC functions.

immunology↗

bioRxiv2024

KIRA6 is an effective and versatile mast cell inhibitor of IgE-mediated activation

Incidents of IgE-mediated, mast cell (MC)-driven allergic diseases are constantly rising and there is an urgent need for the development of novel pharmacological MC stabilizers. Allergen/antigen (Ag)-triggered activation of MCs via crosslinking of the high-affinity receptor for IgE (Fc{varepsilon}RI) is regulated, amongst others, by the coordinated action of various cytosolic tyrosine kinases of the SRC family, e.g. LYN and FYN, which exert positive as well as negative functions. We report that KIRA6, an inhibitor developed for the endoplasmic reticulum (ER) stress sensor IRE1, suppresses IgE-mediated pro-inflammatory MC activation by inhibiting both LYN and FYN. KIRA6 dose-dependently and effectively attenuates Ag-stimulated early signaling (e.g. substrate tyrosine phosphorylation, Ca2+ mobilization, and activation of MAPK pathways) as well as effector functions such as degranulation and pro-inflammatory cytokine production/secretion in murine bone marrow-derived MCs (BMMCs). Moreover, Ag-triggered bronchoconstriction in an ex vivo model of precision-cut lung slices (PCLS), and IgE-mediated stimulation of human MCs were repressed by KIRA6. To get in-depth inside into KIRA6 interaction with three MC-relevant tyrosine kinases, LYN, FYN, and KIT, and to elicit the potential of KIRA6 structure to serve as pharmacophore for the development of respective single-, dual-, or triple-specificity inhibitors, we modeled and evaluated the binding of KIRA6 on the three kinases by applying homology modeling and molecular dynamics simulations, as well as MM GBSA calculations. We found that KIRA6 has a high propensity to bind the inactive state of LYN, FYN, and KIT with comparable affinities. In conclusion, our data suggest the use of novel inhibitors based on the KIRA6 pharmacophore as effective MC stabilizers to improve treatment of pro-inflammatory diseases with MC involvement in need of effective pharmacological interventions.

immunology↗