Pharmacological and genetic reappraisals of protease and oxidative stress pathways in a mouse model of obstructive lung diseases

Shuto, Tsuyoshi; Kamei, Satoshi; Fujikawa, Haruka; Tasaki, Yukihiro; Sugahara, Takuya; Ono, Tomomi; Matsumoto, Chizuru; Sakaguchi, Yuki; Maruta, Kasumi; Nakashima, Rumi; Kawakami, Taisei; Suico, Mary Ann; Kondo, Yoshitaka; Ishigami, Akihito; Tanaka, Takeo; Tanaka, Ken Ichiro; Watanabe, Hiroshi; Nakagata, Naomi; Uchimura, Kohei; Kitamura, Kenichiro; Li, Jian Dong; Kai, Hirofumi

doi:10.1038/srep39305

Cited by 21 publications

(31 citation statements)

References 65 publications

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“…Total RNA was isolated from cells and mouse lung tissues using the RNAiso Plus Kit (Takara) and cDNA synthesis was performed using the PrimeScript RT Regent Kit (Takara). Real-time quantitative RT-PCR analysis was performed as described previously ( Shuto et al, 2016 ). α-, β-, and γ-ENaC-expression plasmids (pCMV-Tag5A) were produced previously ( Sugahara et al, 2009 ) and used to measure the total amounts of mRNA.…”

Section: Methodsmentioning

confidence: 99%

“…Indeed, the expression and function of ENaC were inversely associated with lung function in CF patients ( Boucher, 2007 ). Moreover, we and others have reported that mice with airway-specific overexpression of βENaC (βENaC-transgenic [Tg] mice) exhibit the critical pulmonary phenotypes of CF, such as mucus obstructive, airway inflammatory, and emphysematous phenotypes ( Mall et al, 2004 , Johannesson et al, 2012 , Shuto et al, 2016 ). These findings imply that CFTR and ENaC are essential ion transporters that control airway fluid homeostasis.…”

Section: Introductionmentioning

confidence: 99%

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Zinc Deficiency via a Splice Switch in Zinc Importer ZIP2/SLC39A2 Causes Cystic Fibrosis-Associated MUC5AC Hypersecretion in Airway Epithelial Cells

et al. 2018

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Airway mucus hyperproduction and fluid imbalance are important hallmarks of cystic fibrosis (CF), the most common life-shortening genetic disorder in Caucasians. Dysregulated expression and/or function of airway ion transporters, including cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC), have been implicated as causes of CF-associated mucus hypersecretory phenotype. However, the contributory roles of other substances and transporters in the regulation of CF airway pathogenesis remain unelucidated. Here, we identified a novel connection between CFTR/ENaC expression and the intracellular Zn2 + concentration in the regulation of MUC5AC, a major secreted mucin that is highly expressed in CF airway. CFTR-defective and ENaC-hyperactive airway epithelial cells specifically and highly expressed a unique, alternative splice isoform of the zinc importer ZIP2/SLC39A2 (ΔC-ZIP2), which lacks the C-terminal domain. Importantly, ΔC-ZIP2 levels correlated inversely with wild-type ZIP2 and intracellular Zn2 + levels. Moreover, the splice switch to ΔC-ZIP2 as well as decreased expression of other ZIPs caused zinc deficiency, which is sufficient for induction of MUC5AC; while ΔC-ZIP2 expression per se induced ENaC expression and function. Thus, our findings demonstrate that the novel splicing switch contributes to CF lung pathology via the novel interplay of CFTR, ENaC, and ZIP2 transporters.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Zinc Deficiency via a Splice Switch in Zinc Importer ZIP2/SLC39A2 Causes Cystic Fibrosis-Associated MUC5AC Hypersecretion in Airway Epithelial Cells

et al. 2018

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“…ENaC overexpression/hyperactivation is an important hallmark in the pathogenesis of many types of lung diseases, such as COPD and CF, [4][5][6] and causes impaired mucociliary clearance in the airway. Improvement of "mucostasis," a complication in patients having defects in mucociliary clearance, by targeting ENaC is now widely accepted in the pulmonary research field.…”

Section: Discussionmentioning

confidence: 99%

“…3) Importantly, hyperactivation of ENaC was correlated with lung dysfunction in patients with cystic fibrosis (CF), 4) a common genetic disorder in Caucasians. Moreover, ENaC were also dysregulated in patients with chronic obstructive pulmonary disease (COPD), 5,6) a lung disease that exhibits inflammatory, emphysematous and mucus hypersecretory phenotypes. Consistently, airway-specific βENaC-transgenic (Tg) mice show mucus obstructive lung phenotypes like CF and COPD, 7,8) implying that constitutive and hyperactive ENaC is being recog-nized as therapeutic targets; however, the agents that suppress ENaC are yet to be clinically available.…”

Section: Introductionmentioning

confidence: 99%

Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation <i>in Vitro</i> and Modulates Downstream Pathogenesis <i>in Vivo</i>

Fujikawa

Kawakami

Nakashima

et al. 2020

Biological & Pharmaceutical Bulletin

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Epithelial sodium channel (ENaC) is an amiloride-sensitive sodium ion channel that is expressed in epithelial tissues. ENaC overexpression and/or hyperactivation in airway epithelial cells cause sodium overabsorption and dysregulated ciliary movement for mucus clearance; however, the agents that suppress constitutive airway ENaC activation are yet to be clinically available. Here, we focused on macrolides, which are widely used antibiotics that have many potential immunomodulatory effects. We examined whether macrolides could modulate constitutive ENaC activity and downstream events that typify cystic fibrosis (CF) and chronic obstructive pulmonary diseases (COPD) in in vitro and in vivo models of ENaC overexpression. Treatment of ENaC-overexpressing human bronchial epithelial cells (β/γENaC-16HBE14o-cells) with three macrolides (erythromycin, clarithromycin, azithromycin) confirmed dose-dependent suppression of ENaC function. For in vivo studies, mice harboring airway specific βENaC overexpression (C57BL/6J-βENaCtransgenic mice) were treated orally with azithromycin, a well-established antimicrobial agent that has been widely prescribed. Azithromycin treatment modulated pulmonary mechanics, emphysematous phenotype and pulmonary dysfunction. Notably, a lower dose (3 mg kg 1) of azithromycin significantly increased forced expiratory volume in 0.1 s (FEV0.1), an inverse indicator of bronchoconstriction. Although not statistically significant, improvement of pulmonary obstructive parameters such as emphysema and lung dysfunction (FEV0.1%) was observed. Our results demonstrate that macrolides directly attenuate constitutive ENaC function in vitro and may be promising for the treatment of obstructive lung diseases with defective mucociliary clearance, possibly by targeting ENaC hyperactivation.

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“…A real-time quantitative reverse transcription (RT)-polymerase chain reaction (PCR) analysis. Quantitative RT-PCR was performed by protocols that were previously reported 37 . After reverse transcription, we performed real-time PCR with iQ5 (Bio-Rad) in a mix containing DNA polymerase and SYBR Green (PR820; Takara Bio, Inc., Shiga, Japan).…”

Section: Methodsmentioning

confidence: 99%

The DsbA-L gene is associated with respiratory function of the elderly via its adiponectin multimeric or antioxidant properties

Oniki

Nakashima

Obata

et al. 2020

Sci Rep

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Oxidative stress and inflammation play a key role in the age-related decline in the respiratory function. Adipokine in relation to the metabolic and inflammatory systems is attracting growing interest in the field of respiratory dysfunction. The present clinical and experimental studies investigated the role of the disulfide bond-forming oxidoreductase Alike protein (DsbA-L) gene, which has antioxidant and adiponectin multimeric (i.e. activation) properties, on the respiratory function of the elderly. We performed a retrospective longitudinal genotype-phenotype relationship analysis of 318 Japanese relatively elderly participants (mean age ± standard deviation: 67.0 ± 5.8 years) during a health screening program and an in vitro DsbA-L knock-down evaluation using 16HBE14o-cells, a commonly evaluated human airway epithelial cell line. The DsbA-L rs1917760 polymorphism was associated with a reduction in the ratio of forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC) and %FEV1 and with the elevation of the prevalence of FEV1/FVC < 70%. We also confirmed that the polymorphism was associated with a decreased respiratory function in relation to a decrease in the ratio of high-molecular-weight adiponectin/total adiponectin (as a marker of adiponectin multimerization) and an increase in the oxidized human serum albumin (as an oxidative stress marker). Furthermore, we clarified that DsbA-L knock-down induced oxidative stress and up-regulated the mucus production in human airway epithelial cells. These findings suggest that the DsbA-L gene may play a role in protecting the respiratory function of the elderly, possibly via increased systemic adiponectin functions secreted from adipocytes or through systemic and/or local pulmonary antioxidant properties. Airway inflammation and an impaired lung function are common in elderly populations, leading to decreased quality of life and increase in morbidity and mortality 1-3. Although exposures to cigarette smoke and other environmental pollutants (e.g. biomass fuel exposure and air pollution) are major risk factors for the development and progression of respiratory dysfunction 4,5 , previous evidence has shown that age-related changes in the respiratory functions are also key factors for respiratory dysfunction 1-3. Aging causes the loss of lung elasticity, decreased respiratory muscles, and a decreased surface area for alveolar gas exchange, leading to an impaired respiratory function 1-3. Furthermore, local and/or systemic oxidative stress as well as inflammation play substantial roles in the age-related decline in the respiratory function 1,2. However, these changes in respiratory pathological features

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Pharmacological and genetic reappraisals of protease and oxidative stress pathways in a mouse model of obstructive lung diseases

Cited by 21 publications

References 65 publications

Zinc Deficiency via a Splice Switch in Zinc Importer ZIP2/SLC39A2 Causes Cystic Fibrosis-Associated MUC5AC Hypersecretion in Airway Epithelial Cells

Zinc Deficiency via a Splice Switch in Zinc Importer ZIP2/SLC39A2 Causes Cystic Fibrosis-Associated MUC5AC Hypersecretion in Airway Epithelial Cells

Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation <i>in Vitro</i> and Modulates Downstream Pathogenesis <i>in Vivo</i>

The DsbA-L gene is associated with respiratory function of the elderly via its adiponectin multimeric or antioxidant properties

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