RhoB is associated with the anti-angiogenic effects of celiac patient transglutaminase 2-targeted autoantibodies.
Celiac patient-derived anti-transglutaminase 2 (TG2) antibodies disturb several steps in angiogenesis, but the detailed molecular basis is not known. Therefore, we here analyzed by microarray technology the expression of a set of genes related to angiogenesis and endothelial cell biology in order to identify factors that could explain our previous data related to vascular biology in the context of celiac disease. To this end, in vitro models using human umbilical vein endothelial cells (HUVECs) or in vivo models of angiogenesis were used. A total of 116 genes were analyzed after treatment with celiac patient autoantibodies against TG2. Compared to treatment with control IgA celiac patient, total IgA induced a consistent expression change of 10 genes, the up-regulation of four and down-regulation of six. Of these genes the up-regulated RhoB was selected for further studies. RhoB expression was found to be up-regulated at both messenger RNA and protein level in response to celiac patient total IgA as well as anti-TG2-specific antibody derived from a celiac patient. Interestingly, down-regulation of RhoB by specific small interfering RNA treatment in endothelial cells could rescue the deranged endothelial length and tubule formation caused by celiac disease autoantibodies. RhoB function is controlled by its post-translational modification by farnesylation. This modification of RhoB required for its correct function can be prevented by the cholesterol lowering drug simvastatin, which was also able to abolish the anti-angiogenic effects of celiac anti-TG2 autoantibodies. Taken together, our results would suggest that RhoB plays a key role in the response of endothelial cells to celiac disease-specific anti-TG2 autoantibodies.
Pediatric Research Center, University of Tampere, Tampere University Hospital, Finn-Medi 3, School of Medicine 33014, University of Tampere, Finland.
Stefania
S
Martucciello
Miha
M
Lavric
Boglarka
B
Toth
Toth
T
Boglarka
Ilma
I
Korponay-Szabo
Cristina
C
Nadalutti
Essi
E
Myrsky
Tiina
T
Rauhavirta
Carla
C
Esposito
Ana-Marija
AM
Sulic
Daniele
D
Sblattero
Roberto
R
Marzari
Markku
M
Mäki
Katri
K
Kaukinen
Katri
K
Lindfors
Sergio
S
Caja
D020533:Angiogenesis Inhibitors / Q000276:immunology / Q000494:pharmacology; D000818:Animals; D001323:Autoantibodies / Q000276:immunology* / Q000494:pharmacology; D002446:Celiac Disease / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D016000:Cluster Analysis; D005260:Female; D019204:GTP-Binding Proteins / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D020868:Gene Silencing; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D034622:RNA Interference; D011503:Transglutaminases / Q000276:immunology*; D020743:rhoB GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
D020533:Angiogenesis Inhibitors; D001323:Autoantibodies; D007070:Immunoglobulin A; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins; D020743:rhoB GTP-Binding Protein