Screening Health-Promoting Compounds for Their Capacity to Induce the Activity of FOXO3

Abstract

Several chemical compounds including natural products have been suggested as being effective against age-related diseases or as beneficial for a healthy life. On the other hand, forkhead box O (FOXO) proteins are emerging as key cellular components associated with extreme human longevity. FOXO proteins are mainly regulated by posttranslational modifications and as these modifications are reversible, activation and inactivation of FOXO are attainable through pharmacological treatment. Here, we questioned whether a panel of compounds with known health-beneficial properties has the capacity to induce the activity of FOXO factors. We show that resveratrol, a phytoalexin present in grapes and other food products, the amide alkaloid piperlongumine found in the fruit of the long pepper, and the plant-derived β-carboline compound harmine induced nuclear translocation of FOXO3. We also show that piperlongumine and harmine but not resveratrol activate FOXO-dependent transcription. We determined the half maximal effective concentration (EC50) values for resveratrol, piperlongumine, and harmine for FOXO translocation, and analyzed their inhibitory impact on chromosomal maintenance 1 (CRM1)-mediated nuclear export and the production of reactive oxygen species (ROS). We also used chemical biology approach and Western blot analysis to explore the underlying molecular mechanisms. We show that harmine, piperlongumine, and resveratrol activate FOXO3 independently of phosphoinositide 3-kinase (PI3K)/AKT signaling and the CRM1-mediated nuclear export. The effect of harmine on FOXO3 activity is at least partially mediated through the inhibition of dual-specificity tyrosine (Y) phosphorylationregulated kinase 1A (DYRK1A) and can be reverted by the inhibition of sirtuins (SIRTs).

Multiplexed cellular profiling identifies an organoselenium compound as an inhibitor of CRM1‐mediated nuclear export

“…We have previously established U2nesRELOC, a screening/counter‐screening system that has been successfully used to identify CRM1 inhibitors 26–29 . Based on this reporter system, we developed U2redNES, a stable cell system capable of monitoring CRM1‐mediated nuclear export by using the HIV‐1 protein Rev 30 with a strong heterologous NES 31 fused to red fluorescent protein (RFP).…”

“…To generate the red fluorescent reporter cell line, we based it on the reporter cell line U2nesRELOC that expresses the construct pRev MAPKK GFP, which has been described earlier 26–29 . pRev MAPKK nesGFP carries the NES from MAPK kinase (MAPKK) cloned in pRev(1.4)‐GFP, sandwiched between the Rev and the GFP coding sequences.…”

“…To generate the red fluorescent reporter cell line, we based it on the reporter cell line U2nesRELOC that expresses the construct pRev MAPKK GFP, which has been described earlier. [26][27][28][29]…”

mTORC2 Is the Major Second Layer Kinase Negatively Regulating FOXO3 Activity

“…Statistics were carried out with SPSS software for calculating the percentage of FOXO translocation [ 35 ] (Jimenez et al, 2021). According to this data, the translocation ratio wasplotted in a graph against the concentration used for each compound in a logarithmic scale to approximate the EC50.…”

Modulating undruggable targets to overcome cancer therapy resistance

“…Compound 24 has also the potential to overcome resistance or increase the sensitivity to HER2 inhibitors in breast cancer, and greatly sensitizes to docetaxel chemotherapy, resulting in tumor regression in vivo (Davies et al, 2012). Very recently, healthpromoting compounds have been assessed for their capacity to induce the activity of FOXO3 and the natural occurring alkaloids harmine and piperlongumine were identified as potent activators of FOXO3 nuclear localization (Jimenez, et al, 2021).…”