PRO-INFLAMMATORY WNT5A GENE TRANSCRIPTION IS REGULATED BY NPD1 IN RPE CELLS DURING OS AS PART OF ITS PROSURVIVAL SIGNALING PATHWAY.
Document Type
Presentation
Start Date
22-10-2010 10:45 AM
End Date
22-10-2010 12:00 PM
Description
NPD1 inhibits the expression of pro-inflammatory genes and enhances the expression of antiapoptotic proteins of the Bcl-2 family (Mukherjee et al. 2004; Lukiw et al. 2005) as part of the prosurvival response elicited by oxidative stress stimuli. These events are integrated to influence the RPE cell fate. We hypothesized that NPD1 signaling regulate the expression of a key set of genes that are effectors in the inflammatory response. To determine if NPD1 was capable of influence gene expression triggered by oxidative stress, we performed a genome wide gene expression assessment using Affymetrix U133 plus 2.0 data on more than three hundred NPD1regulated genes. We compared the expression of more than 70000 human genes on normal and 15-LOX-1 silenced ARPE-19 cells undergoing OS when they were treated with 50nM NPD1. Gene Go analysis of the data showed a network of genes related with the Wnt5a/calcium signaling pathway (p< 0.05) and the possible involvement of Notch. As effector of Notch/Hedgehog, Hes1 is transcriptional modulator of Wnt related gene expression. Hes1 is 2.6 folds up-regulated by OS while NPD1 brought the levels to -3.4 folds of regulation related to the control. Accordingly, Wnt5a was found to follow a similar pattern of expression (OS up-regulated by 2.3 folds and NPD1downregulated by -3.8 folds). This pattern was found to be present at the protein level as well. Moreover, Wnt pathway transcriptional regulators NFATC4 and HNF4A are up-regulated from -9.85 and -1.37 in oxidative stress to -4.79 and 2.45 respectively when cell were treated by NPD1 using Real-Time PCR array targeting human transcription factors. Wnt5a/CamKII is a separate Calcium-dependent pathway of Wnt known to be involved in proinflammatory signaling (Pereira et al, 2008 and Pereira et al, 2009). As target of the global Notch/Hedgehog and Wnt regulation emerged in the network obtained from the gene array data EGR1 nuclear receptor up-regulated one order of magnitude by NPD1 and its regulation was confirmed by Real time PCR. The action of EGR1 was implicated in the regulation observed of VEGF-A and calcipressin. These data all together points to NPD1 targeting a specific set of master genes that underlie the anti-apoptotic and anti-inflammatory NPD1 bioactivity.
Recommended Citation
Calandria, Jorgelina M. and Bazan, N. G., "PRO-INFLAMMATORY WNT5A GENE TRANSCRIPTION IS REGULATED BY NPD1 IN RPE CELLS DURING OS AS PART OF ITS PROSURVIVAL SIGNALING PATHWAY." (2010). Dr. Joseph M. Moerschbaecher, III Graduate Research Day. 6.
https://digitalscholar.lsuhsc.edu/grad_rs/2010/poster1/6
PRO-INFLAMMATORY WNT5A GENE TRANSCRIPTION IS REGULATED BY NPD1 IN RPE CELLS DURING OS AS PART OF ITS PROSURVIVAL SIGNALING PATHWAY.
NPD1 inhibits the expression of pro-inflammatory genes and enhances the expression of antiapoptotic proteins of the Bcl-2 family (Mukherjee et al. 2004; Lukiw et al. 2005) as part of the prosurvival response elicited by oxidative stress stimuli. These events are integrated to influence the RPE cell fate. We hypothesized that NPD1 signaling regulate the expression of a key set of genes that are effectors in the inflammatory response. To determine if NPD1 was capable of influence gene expression triggered by oxidative stress, we performed a genome wide gene expression assessment using Affymetrix U133 plus 2.0 data on more than three hundred NPD1regulated genes. We compared the expression of more than 70000 human genes on normal and 15-LOX-1 silenced ARPE-19 cells undergoing OS when they were treated with 50nM NPD1. Gene Go analysis of the data showed a network of genes related with the Wnt5a/calcium signaling pathway (p< 0.05) and the possible involvement of Notch. As effector of Notch/Hedgehog, Hes1 is transcriptional modulator of Wnt related gene expression. Hes1 is 2.6 folds up-regulated by OS while NPD1 brought the levels to -3.4 folds of regulation related to the control. Accordingly, Wnt5a was found to follow a similar pattern of expression (OS up-regulated by 2.3 folds and NPD1downregulated by -3.8 folds). This pattern was found to be present at the protein level as well. Moreover, Wnt pathway transcriptional regulators NFATC4 and HNF4A are up-regulated from -9.85 and -1.37 in oxidative stress to -4.79 and 2.45 respectively when cell were treated by NPD1 using Real-Time PCR array targeting human transcription factors. Wnt5a/CamKII is a separate Calcium-dependent pathway of Wnt known to be involved in proinflammatory signaling (Pereira et al, 2008 and Pereira et al, 2009). As target of the global Notch/Hedgehog and Wnt regulation emerged in the network obtained from the gene array data EGR1 nuclear receptor up-regulated one order of magnitude by NPD1 and its regulation was confirmed by Real time PCR. The action of EGR1 was implicated in the regulation observed of VEGF-A and calcipressin. These data all together points to NPD1 targeting a specific set of master genes that underlie the anti-apoptotic and anti-inflammatory NPD1 bioactivity.