TY  - JOUR
A1  - Godbole, Amod
A1  - Lyga, Sandra
A1  - Lohse, Martin J.
A1  - Calebiro, Davide
T1  - Internalized TSH receptors en route to the TGN induce local G\(_{S}\)-protein signaling and gene transcription
JF  - Nature Communications
N2  - A new paradigm of G-protein-coupled receptor (GPCR) signaling at intracellular sites has recently emerged, but the underlying mechanisms and functional consequences are insufficiently understood. Here, we show that upon internalization in thyroid cells, endogenous TSH receptors traffic retrogradely to the trans-Golgi network (TGN) and activate endogenous Gs-proteins in the retromer-coated compartment that brings them to the TGN. Receptor internalization is associated with a late cAMP/protein kinase A (PKA) response at the Golgi/TGN. Blocking receptor internalization, inhibiting PKA II/interfering with its Golgi/TGN localization, silencing retromer or disrupting Golgi/TGN organization all impair efficient TSH-dependent cAMP response element binding protein (CREB) phosphorylation. These results suggest that retrograde trafficking to the TGN induces local G\(_{S}\)-protein activation and cAMP/PKA signaling at a critical position near the nucleus, which appears required for efficient CREB phosphorylation and gene transcription. This provides a new mechanism to explain the functional consequences of GPCR signaling at intracellular sites and reveals a critical role for the TGN in GPCR signaling.
KW  - G protein-coupled receptors
KW  - fluorescence imaging
KW  - hormone receptors
KW  - trans-Golgi network
Y1  - 2017
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170375
VL  - 8
IS  - 443
ER  - 
TY  - JOUR
A1  - Weibel, Stephanie
A1  - Basse-Luesebrink, Thomas Christian
A1  - Hess, Michael
A1  - Hofmann, Elisabeth
A1  - Seubert, Carolin
A1  - Langbein-Laugwitz, Johanna
A1  - Gentschev, Ivaylo
A1  - Sturm, Volker Jörg Friedrich
A1  - Ye, Yuxiang
A1  - Kampf, Thomas
A1  - Jakob, Peter Michael
A1  - Szalay, Aladar A.
T1  - Imaging of Intratumoral Inflammation during Oncolytic Virotherapy of Tumors by \(^{19}\)F-Magnetic Resonance Imaging (MRI)
JF  - PLoS ONE
N2  - Background
Oncolytic virotherapy of tumors is an up-coming, promising therapeutic modality of cancer therapy. Unfortunately, non-invasive techniques to evaluate the inflammatory host response to treatment are rare. Here, we evaluate \(^{19}\)F magnetic resonance imaging (MRI) which enables the non-invasive visualization of inflammatory processes in pathological conditions by the use of perfluorocarbon nanoemulsions (PFC) for monitoring of oncolytic virotherapy.

Methodology/Principal Findings
The Vaccinia virus strain GLV-1h68 was used as an oncolytic agent for the treatment of different tumor models. Systemic application of PFC emulsions followed by \(^1H\)/\(^{19}\)F MRI of mock-infected and GLV-1h68-infected tumor-bearing mice revealed a significant accumulation of the \(^{19}\)F signal in the tumor rim of virus-treated mice. Histological examination of tumors confirmed a similar spatial distribution of the \(^{19}\)F signal hot spots and \(CD68^+\)-macrophages. Thereby, the \(CD68^+\)-macrophages encapsulate the GFP-positive viral infection foci. In multiple tumor models, we specifically visualized early inflammatory cell recruitment in Vaccinia virus colonized tumors. Furthermore, we documented that the \(^{19}\)F signal correlated with the extent of viral spreading within tumors.

Conclusions/Significance
These results suggest \(^{19}\)F MRI as a non-invasive methodology to document the tumor-associated host immune response as well as the extent of intratumoral viral replication. Thus, \(^{19}\)F MRI represents a new platform to non-invasively investigate the role of the host immune response for therapeutic outcome of oncolytic virotherapy and individual patient response.
KW  - inflammation
KW  - fluorescence microscopy
KW  - oncolytic viruses
KW  - fluorescence imaging
KW  - macrophages
KW  - magnetic resonance imaging
KW  - histology
KW  - in vivo imaging
Y1  - 2013
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130311
VL  - 8
IS  - 3
ER  -