bims-noxint 2020-11-29 papers

Issue of 2020–11–29
four papers selected by
Laia Caja Puigsubira, Uppsala University

Free Radic Biol Med. 2020 Nov 20. pii: S0891-5849(20)31632-4. [Epub ahead of print]

PDIA1 acts as master organizer of NOX1/NOX4 balance and phenotype response in vascular smooth muscle.

Denise de Castro Fernandes, João Wosniak, Renata Castro Gonçalves, Leonardo Yuji Tanaka, Carolina Gonçalves Fernandes, Daniela Bertolini Zanatta, Ana Barbosa Marcondes de Mattos, Bryan Eric Strauss, Francisco Rafael Martins Laurindo.

Changes in vascular smooth muscle cell (VSMC) phenotype underlie disease pathophysiology and are strongly regulated by NOX NADPH oxidases, with Nox1 favoring synthetic proliferative phenotype and Nox4 supporting differentiation. Growth factor-triggered NOX1 expression/activity strictly depends on the chaperone oxidoreductase protein disulfide isomerase-A1 (PDIA1). Intracellular PDIA1 is required for VSMC migration and cytoskeleton organization, while extracellular PDIA1 fine-tunes cytoskeletal mechanoadaptation and vascular remodeling. We hypothesized that PDIA1 orchestrates NOX1/Nox4 balance and VSMC phenotype. Using an inducible PDIA1 overexpression model in VSMC, we showed that early PDIA1 overexpression (for 24-48h) increased NOX1 expression, hydrogen peroxide steady-state levels and spontaneous VSMC migration distances. Sustained PDIA1 overexpression for 72h and 96h supported high NOX1 levels while also increasing NOX4 expression and, remarkably, switched VSMC phenotype to differentiation. Differentiation was preceded by increased nuclear myocardin and serum response factor-response element activation, with no change in cell viability. Both NOX1 and hydrogen peroxide were necessary for later PDIA1-induced VSMC differentiation. In primary VSMC, PDIA1 knockdown decreased nuclear myocardin and increased the proliferating cell nuclear antigen expression. Newly-developed PDIA1-overexpressing mice (TgPDIA1) exhibited normal general and cardiovascular baseline phenotypes. However, in TgPDIA1 carotids, NOX1 was decreased while NOX4 and calponin expressions were enhanced, indicating overdifferentiation vs. normal carotids. Moreover, in a rabbit overdistension injury model during late vascular repair, PDIA1 silencing impaired VSMC redifferentiation and Nox1/Nox4 balance. Our results suggest a model in which PDIA1 acts as an upstream organizer of Nox1/Nox4 balance and related VSMC phenotype, accounting for baseline differentiation setpoint.

Keywords: NADPH oxidase; NOX; Protein disulfide isomerase; differentiation; oxidants; phenotype; redox; vascular smooth muscle cells; vessel remodeling

DOI: https://doi.org/10.1016/j.freeradbiomed.2020.11.020

Arch Pharm Res. 2020 Nov 26.

NRF2 level is negatively correlated with TGF-β1-induced lung cancer motility and migration via NOX4-ROS signaling.

Dayoung Ryu, Jin-Hee Lee, Mi-Kyoung Kwak.

Transforming growth factor-β1 (TGF-β1) is a multifaceted factor in cancer biology that regulates cell proliferation and migration. Overactivation of nuclear factor erythroid 2-like 2 (NFE2L2; NRF2) in cancers has been associated with facilitated tumor growth and therapy resistance; however, role in cancer migration has not been clearly explained yet. In this study, we investigated the role of NRF2 on TGF-β1-induced cell motility/migration. In NRF2-silenced lung cancer A549 cells, both basal and TGF-β1-inducible cell motility/migration increased compared to those in A549. SMAD transcription activity and phosphorylated SMAD2/3 levels were higher in TGF-β1-treated NRF2-low A549 cells than those in A549. Notably, the levels of reactive oxygen species (ROS) that were elevated by TGF-β1 treatment were higher in the NRF2-low A549 than those in control cells, and treatment with ROS scavenger blocked TGF-β1-induced cell motility. As an underlying molecular link, NADPH oxidase 4 (NOX4) was associated with higher ROS elevation and cell motility of NRF2-low A549. NOX4 and TGF-β1-inducible NOX4 levels were higher in NRF2-low A549 cells than those in A549. Moreover, the pharmacological inhibition of NOX4 blocked the TGF-β1-induced motility of NRF2-low A549 cells. Collectively, these results indicate that TGF-β1-induced cell motility/migration is facilitated in NRF2-inhibited lung cancer cells and that high levels of NOX4/ROS are associated with enhanced motility/migration.

Keywords: Cancer metastasis; EMT; Lung cancer; NFE2L2/NRF2; NOX4; TGF-β1

DOI: https://doi.org/10.1007/s12272-020-01298-z

Ann Geriatr Med Res. 2020 Nov 24.

Sestrin2 Attenuates Cellular Senescence by Inhibiting NADPH Oxidase 4 Expression.

Chae Young Hwang, Ying-Hao Han, Seung-Min Lee, Sang-Mi Cho, Dae-Yeul Yu, Ki-Sun Kwon.

Background: Sestrin2 (Sesn2) is involved in the maintenance of metabolic homeostasis and aging via modulation of the 5' AMP-activated protein kinase-mammalian target of rapamycin (AMPK-mTOR) pathway.
Methods: Wild-type and Sesn2 knockout (KO) mice of the 129/SvJ background were maintained in a pathogen-free authorized facility under a 12-hour dark/light cycle at 20°C-22°C and 50%-60% humidity. Mouse embryonic fibroblasts (MEFs) were prepared from 13.5-day-old embryos derived from Sesn2-KO mice mated with each other.
Results: The MEFs from Sesn2-KO mice showed enlarged and flattened morphologies and senescence-associated β-galactosidase activity, accompanied by an elevated level of reactive oxygen species. These senescence phenotypes recovered following treatment with N-acetyl-cysteine. Notably, the mRNA levels of NADPH oxidase 4 (NOX4) and transforming growth factor (TGF)-β were markedly increased in Sesn2-KO MEFs. Treatment of Sesn2-KO MEFs with the NOX inhibitor diphenyleneiodonium and the TGF-β inhibitor SB431542 restored cell growth inhibited by Sesn2-KO.
Conclusion: Sesn2 attenuates cellular senescence via suppression of TGF-β- and NOX4-induced reactive oxygen species generation and subsequent inhibition of AMPK.

Keywords: NOX4; Reactive oxygen species; Senescence; Sestrin2

DOI: https://doi.org/10.4235/agmr.20.0051

Cancer Res. 2020 Nov 23. pii: canres.0622.2020. [Epub ahead of print]

Glutamine-directed migration of cancer-activated fibroblasts facilitates epithelial tumor invasion.

Aida Mestre-Farrera, Marina Bruch-Oms, Raúl Peña, José Rodríguez-Morató, Lorena Alba-Castellón, Laura Comerma, Miguel Quintela-Fandino, Mireia Duñach, Josep Baulida, Óscar J Pozo, Antonio García de Herreros.

Tumors are complex tissues composed of transformed epithelial cells as well as cancer-activated fibroblasts (CAF) that facilitate epithelial tumor cell invasion. We show here that CAF and other mesenchymal cells rely much more on glutamine than epithelial tumor cells; consequently, they are more sensitive to inhibition of glutaminase. Glutamine dependence drove CAF migration towards this amino acid when cultured in low glutamine conditions. CAF also invaded a Matrigel matrix following a glutamine concentration gradient and enhanced the invasion of tumor cells when both cells were co-cultured. Accordingly, glutamine directed invasion of xenografted tumors in immunocompromised mice. Stimulation of glutamine-driven epithelial tumor invasion by fibroblasts required previous CAF activation which involved the TGFb/Snail1 signaling axis. CAF migration towards Gln presented a polarized Akt2 distribution that was modulated by the Gln-dependent activity of TRAF6 and p62 in the migrating front, and depletion of these proteins prevented Akt2 polarization and Gln-driven CAF invasion. Our results demonstrate that glutamine deprivation promotes CAF migration and invasion, which in turn facilitates the movement of tumor epithelial cells towards nutrient-rich territories. These results provide a novel molecular mechanism for how metabolic stress enhances invasion and metastasis.

DOI: https://doi.org/10.1158/0008-5472.CAN-20-0622