{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T10:38:11Z","timestamp":1780396691950,"version":"3.54.1"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2020,11,30]],"date-time":"2020-11-30T00:00:00Z","timestamp":1606694400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:p>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 (TNF\u03b1) 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-, \u03b2-catenin\/Wnt-, mTOR- signaling pathways (SL327, XAV939, or Torin 2). PcdhgC3 KO cells responded stronger to OGD and TNF\u03b1 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.<\/jats:p>","DOI":"10.3389\/fphar.2020.590144","type":"journal-article","created":{"date-parts":[[2020,11,30]],"date-time":"2020-11-30T06:33:55Z","timestamp":1606718035000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":19,"title":["Deletion of Protocadherin Gamma C3 Induces Phenotypic and Functional Changes in Brain Microvascular Endothelial Cells In Vitro"],"prefix":"10.3389","volume":"11","author":[{"given":"Lydia","family":"Gabbert","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina","family":"Dilling","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Meybohm","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malgorzata","family":"Burek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2020,11,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"437","DOI":"10.1007\/s10545-013-9608-0","article-title":"Blood-brain barrier structure and function and the challenges for CNS drug delivery","volume":"36","author":"Abbott","year":"2013","journal-title":"J. Inherit. Metab. Dis."},{"key":"B2","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.nbd.2009.07.030","article-title":"Structure and function of the blood-brain barrier","volume":"37","author":"Abbott","year":"2010","journal-title":"Neurobiol. Dis."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1940","DOI":"10.1177\/0271678x18768443","article-title":"Gelatinolytic activity of autocrine matrix metalloproteinase-9 leads to endothelial de-arrangement in Moyamoya disease","volume":"38","author":"Blecharz-Lang","year":"","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"B4","doi-asserted-by":"publisher","first-page":"631","DOI":"10.1007\/s12975-018-0614-2","article-title":"Interleukin 6-mediated endothelial barrier disturbances can Be attenuated by blockade of the IL6 receptor expressed in brain microvascular endothelial cells","volume":"9","author":"Blecharz-Lang","year":"","journal-title":"Transl. Stroke Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"e4022","DOI":"10.3791\/4022","article-title":"Generation of an immortalized murine brain microvascular endothelial cell line as an in vitro blood brain barrier model","author":"Burek","year":"2012","journal-title":"J. Vis. Exp."},{"key":"B6","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.mce.2008.09.041","article-title":"Cloning and characterization of the murine claudin-5 promoter","volume":"298","author":"Burek","year":"2009","journal-title":"Mol. Cell. Endocrinol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"672","DOI":"10.1007\/s12975-018-0683-2","article-title":"Hypoxia-Induced MicroRNA-212\/132 alter blood-brain barrier integrity through inhibition of tight junction-associated proteins in human and mouse brain microvascular endothelial cells","volume":"10","author":"Burek","year":"2019","journal-title":"Transl. Stroke Res."},{"key":"B8","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1016\/j.neuron.2012.06.039","article-title":"Functional significance of isoform diversification in the protocadherin gamma gene cluster","volume":"75","author":"Chen","year":"2012","journal-title":"Neuron"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1285","DOI":"10.1089\/ars.2011.3929","article-title":"Tight junctions in brain barriers during central nervous system inflammation","volume":"15","author":"Coisne","year":"2011","journal-title":"Antioxid. Redox Signal."},{"key":"B10","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/s12987-020-00192-6","article-title":"Serum-derived factors of breast cancer patients with brain metastases alter permeability of a human blood-brain barrier model","volume":"17","author":"Curtaz","year":"2020","journal-title":"Fluids Barriers CNS"},{"key":"B11","doi-asserted-by":"publisher","first-page":"e1000745","DOI":"10.1371\/journal.pgen.1000745","article-title":"Frequent long-range epigenetic silencing of protocadherin gene clusters on chromosome 5q31 in Wilms\u2019 tumor","volume":"5","author":"Dallosso","year":"2009","journal-title":"PLoS Genet."},{"key":"B12","doi-asserted-by":"publisher","first-page":"4409","DOI":"10.1038\/onc.2011.609","article-title":"Long-range epigenetic silencing of chromosome 5q31 protocadherins is involved in early and late stages of colorectal tumorigenesis through modulation of oncogenic pathways","volume":"31","author":"Dallosso","year":"2012","journal-title":"Oncogene"},{"key":"B13","doi-asserted-by":"publisher","first-page":"648","DOI":"10.1002\/ana.23648","article-title":"The blood-brain barrier in health and disease","volume":"72","author":"Daneman","year":"2012","journal-title":"Ann. Neurol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1038\/nm890","article-title":"RAGE mediates amyloid-\u03b2 peptide transport across the blood-brain barrier and accumulation in brain","volume":"9","author":"Deane","year":"2003","journal-title":"Nat. Med."},{"key":"B15","doi-asserted-by":"publisher","first-page":"3391","DOI":"10.1177\/0271678x16688706","article-title":"Multiple protocadherins are expressed in brain microvascular endothelial cells and might play a role in tight junction protein regulation","volume":"37","author":"Dilling","year":"2017","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"B16","doi-asserted-by":"publisher","first-page":"4074","DOI":"10.4049\/jimmunol.1600823","article-title":"Endothelial MAPKs direct ICAM-1 signaling to divergent inflammatory functions","volume":"198","author":"Dragoni","year":"2017","journal-title":"J. Immunol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1174","DOI":"10.1128\/mcb.00903-10","article-title":"Neuropilin-1 signaling through p130Cas tyrosine phosphorylation is essential for growth factor-dependent migration of glioma and endothelial cells","volume":"31","author":"Evans","year":"2011","journal-title":"Mol. Cell Biol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1007\/s10456-019-09683-z","article-title":"The protein tyrosine phosphatase PTPRJ\/DEP-1 contributes to the regulation of the Notch-signaling pathway and sprouting angiogenesis","volume":"23","author":"Fournier","year":"2020","journal-title":"Angiogenesis"},{"key":"B19","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1016\/j.mcn.2005.05.001","article-title":"Differential expression of individual gamma-protocadherins during mouse brain development","volume":"29","author":"Frank","year":"2005","journal-title":"Mol. Cell. Neurosci."},{"key":"B20","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1016\/j.neuron.2012.01.028","article-title":"\u03b3-Protocadherins control cortical dendrite arborization by regulating the activity of a FAK\/PKC\/MARCKS signaling pathway","volume":"74","author":"Garrett","year":"2012","journal-title":"Neuron"},{"key":"B21","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1002\/ana.25458","article-title":"Plasminogen activator inhibitor\u20101 augments damage by impairing fibrinolysis after traumatic brain injury","volume":"85","author":"Griemert","year":"2019","journal-title":"Ann. Neurol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"9313","DOI":"10.1074\/jbc.m412909200","article-title":"Presenilin-dependent processing and nuclear function of -protocadherins","volume":"280","author":"Haas","year":"2005","journal-title":"J. Biol. Chem."},{"key":"B23","doi-asserted-by":"publisher","first-page":"2217","DOI":"10.1161\/atvbaha.118.311321","article-title":"Direct targeting of the mTOR (mammalian target of rapamycin) kinase improves endothelial permeability in drug-eluting stents-brief report","volume":"38","author":"Harari","year":"2018","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"862","DOI":"10.1177\/0271678x16630991","article-title":"In vitro models of the blood-brain barrier: an overview of commonly used brain endothelial cell culture models and guidelines for their use","volume":"36","author":"Helms","year":"2016","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"B25","doi-asserted-by":"publisher","first-page":"83","DOI":"10.3389\/fneur.2019.00083","article-title":"IL-6 plasma levels correlate with cerebral perfusion deficits and infarct sizes in stroke patients without associated infections","volume":"10","author":"Hotter","year":"2019","journal-title":"Front. Neurol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"73","DOI":"10.3389\/fcvm.2020.00073","article-title":"Increased catecholamine levels and inflammatory mediators alter barrier properties of brain microvascular endothelial cells in vitro","volume":"7","author":"Ittner","year":"2020","journal-title":"Front. Cardiovasc. Med."},{"key":"B27","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1177\/1073858402008003012","article-title":"Interleukin-6 (IL-6): a possible neuromodulator induced by neuronal activity","volume":"8","author":"J\u00fcttler","year":"2002","journal-title":"Neuroscientist"},{"key":"B28","doi-asserted-by":"publisher","first-page":"122","DOI":"10.3390\/ijms20010122","article-title":"The influence of capsaicin on the integrity of microvascular endothelial cell monolayers","volume":"20","author":"Kaiser","year":"2018","journal-title":"Int. J. Mol. Sci."},{"key":"B29","doi-asserted-by":"publisher","first-page":"30551","DOI":"10.1074\/jbc.m605677200","article-title":"Allelic gene regulation ofPcdh-\u03b1 andPcdh-\u03b3 clusters involving both monoallelic and biallelic expression in single purkinje cells","volume":"281","author":"Kaneko","year":"2006","journal-title":"J. Biol. Chem."},{"key":"B30","doi-asserted-by":"publisher","first-page":"613","DOI":"10.1146\/annurev-pharmtox-010814-124852","article-title":"Targeting receptor-mediated transport for delivery of biologics across the blood-brain barrier","volume":"55","author":"Lajoie","year":"2015","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1083\/jcb.200209019","article-title":"Contact inhibition of VEGF-induced proliferation requires vascular endothelial cadherin, \u03b2-catenin, and the phosphatase DEP-1\/CD148","volume":"161","author":"Lampugnani","year":"2003","journal-title":"J. Cell Biol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"737","DOI":"10.1038\/376737a0","article-title":"Protein tyrosine kinase PYK2 involved in Ca2+-induced regulation of ion channel and MAP kinase functions","volume":"376","author":"Lev","year":"1995","journal-title":"Nature"},{"key":"B33","doi-asserted-by":"publisher","first-page":"913","DOI":"10.1111\/j.1471-4159.2011.07587.x","article-title":"Molecular heterogeneity in the choroid plexus epithelium: the 22-member \u03b3-protocadherin family is differentially expressed, apically localized, and implicated in CSF regulation","volume":"120","author":"Lobas","year":"2012","journal-title":"J. Neurochem."},{"key":"B34","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1186\/1742-2094-7-14","article-title":"Cerebral ischemia induces micro vascular pro-inflammatory cytokine expression via the MEK\/ERK pathway","volume":"7","author":"Maddahi","year":"2010","journal-title":"J. Neuroinflammation"},{"key":"B35","doi-asserted-by":"publisher","first-page":"31665","DOI":"10.1038\/srep31665","article-title":"The gamma-Protocadherin-C3 isoform inhibits canonical Wnt signalling by binding to and stabilizing Axin1 at the membrane","volume":"6","author":"Mah","year":"2016","journal-title":"Sci. Rep."},{"key":"B36","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.semcdb.2017.07.043","article-title":"Regulation of Wnt signaling by protocadherins","volume":"69","author":"Mah","year":"2017","journal-title":"Semin. Cell Dev. Biol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"840","DOI":"10.1002\/cne.24783","article-title":"Expression of protocadherin\u2010\u03b3C4 protein in the rat brain","volume":"528","author":"Miralles","year":"2020","journal-title":"J. Comp. Neurol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"2702","DOI":"10.1016\/j.celrep.2017.02.060","article-title":"\u03b3-Protocadherins interact with neuroligin-1 and negatively regulate dendritic spine morphogenesis","volume":"18","author":"Molumby","year":"2017","journal-title":"Cell Rep."},{"key":"B39","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1006\/jmbi.2000.3777","article-title":"Phylogenetic analysis of the cadherin superfamily allows identification of six major subfamilies besides several solitary members 1 1Edited by M. Yaniv","volume":"299","author":"Nollet","year":"2000","journal-title":"J. Mol. Biol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.1093\/carcin\/23.7.1139","article-title":"Protocadherin LKC, a new candidate for a tumor suppressor of colon and liver cancers, its association with contact inhibition of cell proliferation","volume":"23","author":"Okazaki","year":"2002","journal-title":"Carcinogenesis"},{"key":"B41","doi-asserted-by":"publisher","first-page":"4133","DOI":"10.1007\/s00018-017-2572-3","article-title":"Regulation of neural circuit formation by protocadherins","volume":"74","author":"Peek","year":"2017","journal-title":"Cell. Mol. Life Sci."},{"key":"B42","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.semcdb.2017.05.001","article-title":"Clustered protocadherin trafficking","volume":"69","author":"Phillips","year":"2017","journal-title":"Semin. Cell Dev. Biol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"38527","DOI":"10.1074\/jbc.m104536200","article-title":"Synergistic cooperation between hypoxia and transforming growth factor-\u03b2 pathways on human vascular endothelial growth factor gene expression","volume":"276","author":"S\u00e1nchez-Elsner","year":"2001","journal-title":"J. Biol. Chem."},{"key":"B44","doi-asserted-by":"publisher","first-page":"2249","DOI":"10.1002\/j.1460-2075.1993.tb05878.x","article-title":"Protocadherins: a large family of cadherin-related molecules in central nervous system","volume":"12","author":"Sano","year":"1993","journal-title":"EMBO J."},{"key":"B45","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1016\/0090-1229(89)90010-x","article-title":"Endotoxin, tumor necrosis factor-\u03b1 and interleukin 1 induce interleukin 6 production in vivo","volume":"53","author":"Shalaby","year":"1989","journal-title":"Clin. Immunol. Immunopathol."},{"key":"B46","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.jneuroim.2006.06.019","article-title":"Differential susceptibility of cerebral and cerebellar murine brain microvascular endothelial cells to loss of barrier properties in response to inflammatory stimuli","volume":"179","author":"Silwedel","year":"2006","journal-title":"J. Neuroimmunol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"192","DOI":"10.18632\/aging.100296","article-title":"Roles of the Raf\/MEK\/ERK and PI3K\/PTEN\/Akt\/mTOR pathways in controlling growth and sensitivity to therapy-implications for cancer and aging","volume":"3","author":"Steelman","year":"2011","journal-title":"Aging (Albany NY)"},{"key":"B48","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1093\/jmcb\/mjs034","article-title":"Protocadherin clusters and cell adhesion kinase regulate dendrite complexity through Rho GTPase","volume":"4","author":"Suo","year":"2012","journal-title":"J. Mol. Cell Biol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"3","DOI":"10.3390\/nano6010003","article-title":"Receptor-mediated drug delivery systems targeting to glioma","volume":"6","author":"Wang","year":"2015","journal-title":"Nanomaterials (Basel)"},{"key":"B50","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1016\/j.biopha.2016.11.021","article-title":"Rapamycin inhibits the proliferation of endothelial cells in hemangioma by blocking the mTOR-FABP4 pathway","volume":"85","author":"Wang","year":"2017","journal-title":"Biomed. Pharmacother."},{"key":"B51","doi-asserted-by":"publisher","first-page":"4","DOI":"10.3389\/fnmol.2013.00004","article-title":"Protocadherins, not prototypical: a complex tale of their interactions, expression, and functions","volume":"6","author":"Weiner","year":"2013","journal-title":"Front. Mol. Neurosci."},{"key":"B52","doi-asserted-by":"publisher","first-page":"7","DOI":"10.3389\/fneng.2013.00007","article-title":"The blood-brain barrier: an engineering perspective","volume":"6","author":"Wong","year":"2013","journal-title":"Front. Neuroeng."},{"key":"B53","doi-asserted-by":"publisher","first-page":"1396","DOI":"10.1016\/j.bbrc.2006.02.100","article-title":"Interleukin-6 triggers human cerebral endothelial cells proliferation and migration: the role for KDR and MMP-9","volume":"342","author":"Yao","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B54","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1111\/jcmm.12022","article-title":"Loss of CCM3 impairs DLL4-Notch signalling: implication in endothelial angiogenesis and in inherited cerebral cavernous malformations","volume":"17","author":"You","year":"2013","journal-title":"J. Cell Mol. Med."},{"key":"B55","doi-asserted-by":"publisher","first-page":"4657","DOI":"10.1038\/onc.2008.101","article-title":"PCDH8, the human homolog of PAPC, is a candidate tumor suppressor of breast cancer","volume":"27","author":"Yu","year":"2008","journal-title":"Oncogene"},{"key":"B56","doi-asserted-by":"publisher","first-page":"829","DOI":"10.1172\/jci9369","article-title":"VEGF enhances angiogenesis and promotes blood-brain barrier leakage in the ischemic brain","volume":"106","author":"Zhang","year":"2000","journal-title":"J. Clin. Invest."},{"key":"B57","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1158\/0008-5472.can-16-1267-t","article-title":"PROTOCADHERIN 7 acts through SET and PP2A to potentiate MAPK signaling by EGFR and KRAS during lung tumorigenesis","volume":"77","author":"Zhou","year":"2017","journal-title":"Cancer Res."},{"key":"B58","doi-asserted-by":"publisher","first-page":"1897","DOI":"10.1007\/s11064-018-2607-7","article-title":"TLR2 ligand Pam3CSK4 regulates MMP-2\/9 expression by MAPK\/NF-\u03baB signaling pathways in primary brain microvascular endothelial cells","volume":"43","author":"Zhu","year":"2018","journal-title":"Neurochem. Res."}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2020.590144\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,12,17]],"date-time":"2020-12-17T17:33:17Z","timestamp":1608226397000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2020.590144\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,30]]},"references-count":58,"alternative-id":["10.3389\/fphar.2020.590144"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2020.590144","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,11,30]]},"article-number":"590144"}}