CD40: an upstream master switch for endothelial cell activation uncovered by RNAi-coupled transcriptional profiling.
The CD40-CD154 dyad seems to play a prominent role fostering the immune-inflammatory response triggered by endothelial cell (EC)-T-cell communication. To delineate comprehensively the involvement of CD40 (TNFRSF5) in EC activation, we combined RNAi-mediated CD40 knockdown with comparative genome-wide transcriptional profiling of ECs interacting with (CD154+) T cells. We report the initiation of a profound stress response in ECs upon CD40-CD154 engagement through early up-regulation of, among others, the major proinflammatory NF-kappaB and MAPK/SAPK pathways and their associated transcription factors. Moreover, we have identified novel genes regulated through the CD40-CD154 interaction, and pathways previously unrecognized to be induced by CD40 signaling in ECs. Thus, we document a significant down-regulation of endothelial APLN by CD40-CD154 interaction, TNFalpha/IFNgamma exposure, and in immune-inflammatory pathologies, which could lead to hemodynamic dysfunction. Conversely, CD40-mediated up-regulation of the viral immune surveillance system, notably TLR3, IFIH1, RIG-I, and RNASEL, establishes a reverse link from adaptive to innate immunity in ECs. Moreover, systematic enrichment analysis substantiates endothelial CD40 involvement in the transcriptional regulation of gene networks associated with adhesion and motility, immunity, cell fate control, hemostasis, and metabolism. Our study also highlights the anti-inflammatory potential of RNAi-mediated CD40 inhibition, and the relevance of CD40 signaling for therapeutic intervention.
Pluvinet
Raquel
R
Medical and Molecular Genetics Center, Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), Hospital Duran i Reynals, L'Hospitalet de Llobregat, Barcelona, Spain.
Olivar
Rut
R
Krupinski
Jerzy
J
Herrero-Fresneda
Inmaculada
I
Luque
Anna
A
Torras
Joan
J
Cruzado
Josep M
JM
Grinyó
Josep M
JM
Sumoy
Lauro
L
Aran
Josep M
JM
D000818:Animals; D000073861:Apelin; D019013:CD40 Antigens / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D023201:CD40 Ligand / Q000378:metabolism; D002450:Cell Communication; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000276:immunology* / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D019169:Jurkat Cells; D016030:Kidney Transplantation / Q000276:immunology; D008213:Lymphocyte Activation; D020935:MAP Kinase Signaling System; D016328:NF-kappa B / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D011914:Rats, Inbred BN; D017208:Rats, Wistar; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology
C115436:APLN protein, human; D000073861:Apelin; D019013:CD40 Antigens; D018836:Inflammation Mediators; D036341:Intercellular Signaling Peptides and Proteins; D016328:NF-kappa B; D034741:RNA, Small Interfering; D023201:CD40 Ligand