Specificity of NHERF1 regulation of GPCR signaling and function in human airway smooth muscle

Pera, Tonio; Tompkins, Eric; Katz, Michael; Wang, Bin; Deshpande, Deepak A.; Weinman, Edward J.; Penn, Raymond B.

doi:10.1096/fj.201900323r

Cited by 10 publications

(11 citation statements)

References 57 publications

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“…NHERF1 levels were augmented during LPS-mediated inflammation, and inflammation was reduced when NHERF1 was silenced. These results are consistent with previous reports that have demonstrated NHERF1 knockdowns abrogate LPS-induced cAMP-mediated IL-6 production in human airway smooth cells [ 15 ]. Similarly, inhibition of NHERF1-CXCR2 has been shown to prevent chemotaxis and neutrophil adhesion driven by IL-8 [ 73 ].…”

Section: Discussionsupporting

confidence: 93%

“…For example, NHERF1 is a positive regulator of NF-kB activation in vascular inflammation by recruiting PKCζ at the cell membrane, resulting in activation of NF-kB [ 14 ]. NHERF1 maintains cyclic amino monophosphate (cAMP) and PKA signaling and regulates interleukin (IL)-6 production in human airway smooth muscle cells [ 15 ] and promotes mast cell activation via the C3a complement receptor (C3aR) pathway [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

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Sodium Hydrogen Exchanger Regulatory Factor-1 (NHERF1) Regulates Fetal Membrane Inflammation

Kammala

Sheller‐Miller

Radnaa

et al. 2020

IJMS

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The fetal inflammatory response, a key contributor of infection-associated preterm birth (PTB), is mediated by nuclear factor kappa B (NF-kB) activation. Na+/H+ exchanger regulatory factor-1 (NHERF1) is an adapter protein that can regulate intracellular signal transduction and thus influence NF-kB activation. Accordingly, NHERF1 has been reported to enhance proinflammatory cytokine release and amplify inflammation in a NF-kB-dependent fashion in different cell types. The objective of this study was to examine the role of NHERF1 in regulating fetal membrane inflammation during PTB. We evaluated the levels of NHERF1 in human fetal membranes from term labor (TL), term not in labor (TNIL), and PTB and in a CD1 mouse model of PTB induced by lipopolysaccharide (LPS). Additionally, primary cultures of fetal membrane cells were treated with LPS, and NHERF1 expression and cytokine production were evaluated. Gene silencing methods using small interfering RNA targeting NHERF1 were used to determine the functional relevance of NHERF1 in primary cultures. NHERF1 expression was significantly (p < 0.001) higher in TL and PTB membranes compared to TNIL membranes, and this coincided with enhanced (p < 0.01) interleukin (IL)-6 and IL-8 expression levels. LPS-treated animals delivering PTB had increased levels of NHERF1, IL-6, and IL-8 compared to phosphate-buffered saline (PBS; control) animals. Silencing of NHERF1 expression resulted in a significant reduction in NF-kB activation and IL-6 and IL-8 production as well as increased IL-10 production. In conclusion, downregulation of NHERF1 increased anti-inflammatory IL-10, and reducing NHERF1 expression could be a potential therapeutic strategy to reduce the risk of infection/inflammation associated with PTB.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Sodium Hydrogen Exchanger Regulatory Factor-1 (NHERF1) Regulates Fetal Membrane Inflammation

Kammala

Sheller‐Miller

Radnaa

et al. 2020

IJMS

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“…Human ASM cells were treated with PDGF with or without the test compounds, and after 48 h the cell culture medium was harvested for determining IL-6 secretion by ASM cells. Total IL-6 content in the cell culture media was determined by ELISA (R&D Systems, Minneapolis, MN, USA) as previously described by Pera et al (38) in 96-well plates coated with anti-IL-6 antibody.…”

Section: Il-6 Elisa Assaymentioning

confidence: 99%

Effects of ATP‐competitive and function‐selective ERK inhibitors on airway smooth muscle cell proliferation

et al. 2019

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Increased airway smooth muscle (ASM) cell mass and secretory functions are characteristics of airway inflammatory diseases, such as asthma. To date, there are no effective therapies to combat ASM cell proliferation, which contributes to bronchoconstriction and airway obstruction. Growth factors such as platelet‐derived growth factor (PDGF) and the activation of the ERK1/2 are major regulators of ASM cell proliferation and airway remodeling in asthma. However, given the ubiquitous expression and multiple functions of ERK1/2, complete inhibition of ERK1/2 using ATP‐competitive inhibitors may lead to unwanted off‐target effects. Alternatively, we have identified compounds that are designed to target substrate docking sites and act as function‐selective inhibitors of ERK1/2 signaling. Here, we show that both function‐selective and ATP‐competitive ERK1/2 inhibitors are effective at inhibiting PDGF‐mediated proliferation, collagen production, and IL‐6 secretion in ASM cells. Proteomic analysis revealed that both types of inhibitors had similar effects on reducing proteins related to TGF‐β and IL‐6 signaling that are relevant to airway remodeling. However, function‐selective ERK1/2 inhibitors caused fewer changes in protein expression compared with ATP‐competitive inhibitors. These studies provide a molecular basis for the development of function‐selective ERK1/2 inhibitors to mitigate airway remodeling in asthma with defined regulation of ERK1/2 signaling.—Defnet, A. E., Huang, W., Polischak, S., Yadav, S. K., Kane, M. A., Shapiro, P., Deshpande, D. A. Effects of ATP‐competitive and function‐selective ERK inhibitors on airway smooth muscle cell proliferation. FASEB J. 33, 10833–10843 (2019). http://www.fasebj.org

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“…A full evaluation of the alterations in proximal tubule cell transport in the NHERF1 KO kidneys has not been examined. The absence of NHERF1 may also result in impaired signaling processes [53], alterations in intracellular or mitochondrial protein phosphorylation, loss of mitochondrial interaction with other organelles, or aberrant mitochondrial protein localization, as suggested by the proteomic data demonstrating changes in mitochondrial proteins associated with the renal cortical BBM ( Figure S1) (Table S1). Thus, further studies to compare mitochondrial function of WT and NHERF1 KO in intact kidney tissue are warranted.…”

Section: Discussionmentioning

confidence: 99%

NHERF1 Loss Upregulates Enzymes of the Pentose Phosphate Pathway in Kidney Cortex

et al. 2020

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(1) Background: We previously showed Na/H exchange regulatory factor 1 (NHERF1) loss resulted in increased susceptibility to cisplatin nephrotoxicity. NHERF1-deficient cultured proximal tubule cells and proximal tubules from NHERF1 knockout (KO) mice exhibit altered mitochondrial protein expression and poor survival. We hypothesized that NHERF1 loss results in changes in metabolic pathways and/or mitochondrial dysfunction, leading to increased sensitivity to cisplatin nephrotoxicity. (2) Methods: Two to 4-month-old male wildtype (WT) and KO mice were treated with vehicle or cisplatin (20 mg/kg dose IP). After 72 h, kidney cortex homogenates were utilized for metabolic enzyme activities. Non-treated kidneys were used to isolate mitochondria for mitochondrial respiration via the Seahorse XF24 analyzer. Non-treated kidneys were also used for LC-MS analysis to evaluate kidney ATP abundance, and electron microscopy (EM) was utilized to evaluate mitochondrial morphology and number. (3) Results: KO mouse kidneys exhibit significant increases in malic enzyme and glucose-6 phosphate dehydrogenase activity under baseline conditions but in no other gluconeogenic or glycolytic enzymes. NHERF1 loss does not decrease kidney ATP content. Mitochondrial morphology, number, and area appeared normal. Isolated mitochondria function was similar between WT and KO. Conclusions: KO kidneys experience a shift in metabolism to the pentose phosphate pathway, which may sensitize them to the oxidative stress imposed by cisplatin.

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Specificity of NHERF1 regulation of GPCR signaling and function in human airway smooth muscle

Cited by 10 publications

References 57 publications

Sodium Hydrogen Exchanger Regulatory Factor-1 (NHERF1) Regulates Fetal Membrane Inflammation

Sodium Hydrogen Exchanger Regulatory Factor-1 (NHERF1) Regulates Fetal Membrane Inflammation

Effects of ATP‐competitive and function‐selective ERK inhibitors on airway smooth muscle cell proliferation

NHERF1 Loss Upregulates Enzymes of the Pentose Phosphate Pathway in Kidney Cortex

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