TY  - JOUR
A1  - Gabbert, Lydia
A1  - Dilling, Christina
A1  - Meybohm, Patrick
A1  - Burek, Malgorzata
T1  - Deletion of Protocadherin Gamma C3 Induces Phenotypic and Functional Changes in Brain Microvascular Endothelial Cells In Vitro
JF  - Frontiers in Pharmacology
N2  - Inflammation of the central nervous system (CNS) is associated with diseases such as multiple sclerosis, stroke and neurodegenerative diseases. Compromised integrity of the blood-brain barrier (BBB) and increased migration of immune cells into the CNS are the main characteristics of brain inflammation. Clustered protocadherins (Pcdhs) belong to a large family of cadherin-related molecules. Pcdhs are highly expressed in the CNS in neurons, astrocytes, pericytes and epithelial cells of the choroid plexus and, as we have recently demonstrated, in brain microvascular endothelial cells (BMECs). Knockout of a member of the Pcdh subfamily, PcdhgC3, resulted in significant changes in the barrier integrity of BMECs. Here we characterized the endothelial PcdhgC3 knockout (KO) cells using paracellular permeability measurements, proliferation assay, wound healing assay, inhibition of signaling pathways, oxygen/glucose deprivation (OGD) and a pro-inflammatory cytokine tumor necrosis factor alpha (TNFα) treatment. PcdhgC3 KO showed an increased paracellular permeability, a faster proliferation rate, an altered expression of efflux pumps, transporters, cellular receptors, signaling and inflammatory molecules. Serum starvation led to significantly higher phosphorylation of extracellular signal-regulated kinases (Erk) in KO cells, while no changes in phosphorylated Akt kinase levels were found. PcdhgC3 KO cells migrated faster in the wound healing assay and this migration was significantly inhibited by respective inhibitors of the MAPK-, β-catenin/Wnt-, mTOR- signaling pathways (SL327, XAV939, or Torin 2). PcdhgC3 KO cells responded stronger to OGD and TNFα by significantly higher induction of interleukin 6 mRNA than wild type cells. These results suggest that PcdhgC3 is involved in the regulation of major signaling pathways and the inflammatory response of BMECs.
KW  - blood-brain barrier
KW  - protocadherin gamma C3
KW  - inflammation
KW  - oxygen/glucose deprivation
KW  - stroke
KW  - tumor necrosis factor-α
KW  - proliferation
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-219828
SN  - 1663-9812
VL  - 11
ER  - 
TY  - THES
A1  - Gabbert, Lydia
T1  - Protocadherine an der Blut-Hirn-Schranke
T1  - Protocadherins at the blood-brain-barrier
N2  - Protocadherine (Pcdh) sind im zentralen Nervensystem (ZNS) stark exprimiert und üben vielfältige Funktionen bei der neuronalen Entwicklung aus. Der Knockout eines Vertreters der Pcdhs, PcdhgC3, führt zu Veränderungen in tight junction Proteinleveln in mikrovaskulären Endothelzellen der Blut-Hirn-Schranke (BBB). In dieser Arbeit untersuche ich die Rolle des PcdhgC3 Knockouts (KO) in Transportern der Blut-Hirn-Schranke sowie dessen Auswirkungen auf die Signaltransduktion mittels Serumreduktion, Zellmigrationsversuchen, Signalweg-Inhibierung und Sauerstoff-Glucose-Entzug. Der PcdhgC3 Knockout resultiert in veränderten Proteinleveln der BBB Transporter und könnte ein vielversprechendes Therapieziel zukünftiger Pharmakotherapie sein. Ebenso führt die Serumreduktion in den KO-Zellen zu höheren Leveln von Signalkinasen (Erk). Die Knockout-Zelllinie zeigt signifikant schnellere Migrationsraten und scheint durch Signalweg-Inhibitoren (mTOR, MAPK, wnt-Inhibitoren) stärker im Wachstum reduziert zu sein. So könnte PcdhgC3 eine Rolle bei der Regulierung von Signalwegen spielen und zu einer veränderten Integrität der Blut-Hirn-Schranke beitragen.
N2  - Protocadherins (Pcdh) are highly expressed in the central nervous system (CNS) and play multiple roles in neuronal development. Knockout of one member of the Pcdh family, PcdhgC3, resulted in changes of tight junction proteins in microvascular endothelial cells of the blood-brain-barrier (BBB). Here I characterize the role of PcdhgC3 knockout (KO) in BBB transporters and its impact on signal transduction using serum starvation, wound healing assays, inhibition of signalling pathways and oxygen glucose deprivation (OGD) tests. PcdhgC3 Knockout resulted in changes in BBB transporter protein levels and might be a promising target for future drug therapy. Serum starvation led to higher levels of signal regulated kinases (Erk) in Knockout cells. KO cells show significant faster migration rates in wound healing assays and are inhibited by signalling pathway inhibitors (mTOR, MAPK, wnt inhibitors) suggesting that PcdhgC3 may play a role in the regulation of signalling pathways and changes of the BBB integrity.
KW  - Cadherine
KW  - Blut-Hirn-Schranke
KW  - Protocadherine
KW  - PcdhgC3
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-234807
ER  -