Differential estrogen‐receptor activation regulates extracellular matrix deposition in human airway smooth muscle remodeling via NF‐κB pathway

Abstract

Altered airway smooth muscle (ASM) mass and extracellular matrix (ECM) deposition in airways are characteristic features of remodeling in asthma. Increased ECM production modulates ASM cell proliferation and leads to airway remodeling. Our previous studies showed that ASM from patients with asthma exhibited increased expression of estrogen receptor (ER)-b, which upon activation down-regulated ASM proliferation, implicating an important role for estrogen signaling in airway physiology. There is no current information on the effect of differential ER activation on ECM production. In this study, we evaluated the effect of ER-a vs. ER-b activation on ECM production, deposition, and underlying pathways. Primary human ASM cells isolated from asthmatics and nonasthmatics were treated with E 2 , an ER-a agonist [propylpyrazoletriol (PPT)], and an ER-b agonist [WAY-200070 (WAY)] with TNF-a or platelet-derived growth factor (PDGF) followed by evaluation of ECM production and deposition. Expression of proteins and genes corresponding to ECM were measured using Western blotting and quantitative RT-PCR with subsequent matrix metalloproteinase (MMP) activity. Molecular mechanisms of ER activation in regulating ECM were evaluated by luciferase reporter assays for activator protein 1 (AP-1) and NF-kB. TNF-a or PDGF significantly (P < 0.001) increased ECM deposition and MMP activity in human ASM cells, which was significantly reduced with WAY treatment but not with PPT. Furthermore, TNF-a-or PDGF-induced ECM gene expression in ASM cells was significantly reduced with WAY (P < 0.001). Moreover, WAY significantly down-regulated the activation of NF-kB (P < 0.001) and AP-1 (P < 0.01, P < 0.05) in ASM cells from asthmatics and nonasthmatics. Overall, we demonstrate differential ER signaling in controlling ECM production and deposition. Activation of ER-b diminishes ECM deposition via suppressing the NF-kB pathway activity and might serve as a novel target to blunt airway remodeling.-

Sex steroids skew ACE2 expression in human airway: a contributing factor to sex differences in COVID-19?

“…Perhaps more interestingly, Tes exposure significantly upregulated ACE2 expression in human ASM cells. Here, it is important to note that E 2 and Tes concentrations used in our study are physiological ( 4 , 5 , 7 , 22 , 38 , 39 ). Furthermore, we have previously reported that both male and female human ASM expresses estrogen receptor isoforms (alpha and beta) as well as the androgen receptor ( 6 , 22 ).…”

“…Serum-deprived human ASM cells were exposed to vehicle, E 2 [1 nM ( 5 , 7 )], and Tes (10 nM ( 22 ),) in DMEM/F12 (FBS free) for 24 h followed by protein collection for Western analyses ( 4 , 5 , 22 ).…”

Role of Estrogen Receptors α and β in a Murine Model of Asthma: Exacerbated Airway Hyperresponsiveness and Remodeling in ERβ Knockout Mice

“…Furthermore, we have also shown that ERb activation inhibits PDGF induced proliferation in primary human ASM cells via AKT/ERK/p38 pathways (Ambhore et al, 2018). Many studies have shown the impact of ER signaling on asthma in vitro (Townsend et al, 2010;Townsend et al, 2012b;Martin et al, 2015;Sathish et al, 2015a;Aravamudan et al, 2017;Ambhore et al, 2018;Ambhore et al, 2019b;Bhallamudi et al, 2019); however, very few studies have explored the role of estrogen in asthma in vivo (Carey et al, 2007a;Carey et al, 2007c;Riffo-Vasquez et al, 2007;Dimitropoulou et al, 2009;Jia et al, 2011). Few studies reported down-regulated AHR upon administration of estrogen in females and OVX mice; however, the receptorspecific role of estrogen has not been explored (Riffo-Vasquez et al, 2007;Matsubara et al, 2008;Dimitropoulou et al, 2009).…”

“…Incidence of asthma is more common in pre-pubescent boys and adult women and the severity of asthma is increased during pregnancy (de Marco et al, 2000;Caracta, 2003;Carey et al, 2007b;Bonds and Midoro-Horiuti, 2013). In this context, multiple studies have explored and suggested a role for sex hormones in airway biology, especially estrogen (Keselman and Heller, 2015;Sathish et al, 2015a;Ambhore et al, 2018;Ambhore et al, 2019a;Ambhore et al, 2019b;Bhallamudi et al, 2019;Fuentes et al, 2019;Fuentes and Silveyra, 2019a). However, there is still a debate on the contradicting role of estrogen as few studies suggest its role in reducing inflammation (Myers and Sherman, 1994;Lieberman et al, 1995;Haggerty et al, 2003), while others suggest estrogen to induce AHR and inflammation (Riffo-Vasquez et al, 2007;Sakazaki et al, 2008).…”

“…Estrogen primarily acts through estrogen receptors (ER) ERa and ERb, which are widely considered to be nuclear receptors (Altucci and Gronemeyer, 2001). Multiple studies suggest both ERa and ERb signal through different cell signaling pathways eliciting different effects on cellular functions (Kuiper et al, 1996;Clarke et al, 2003;Heldring et al, 2007;Edvardsson et al, 2011;Williams and Lin, 2013;Aravamudan et al, 2017;Ambhore et al, 2018;Ambhore et al, 2019b;Bhallamudi et al, 2019;Fuentes and Silveyra, 2019a). ER's are expressed on a wide array of cells and our group has recently showed differential ER expression in human ASM cells, which is upregulated during inflammation/ asthma (especially ERb) (Aravamudan et al, 2017).…”