Targeting the Intracellular Environment in Cystic Fibrosis: Restoring Autophagy as a Novel Strategy to Circumvent the CFTR Defect

Villella, Valeria Rachela; Esposito, Speranza; Bruscia, Emanuela M.; Maiuri, Maria Chiara; Raia, Valeria; Kroemer, Guido; Maiuri, Luigi

doi:10.3389/fphar.2013.00001

Cited by 116 publications

(151 citation statements)

References 118 publications

(159 reference statements)

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“…Driven by the positive outcome of the mouse experiments, we next investigated the possibility that cysteamine, like cystamine, 28,31,40 would rescue F508del-CFTR at the plasma membrane (PM) and sustain the residence of CFTR mutant after washout in vitro, in polarized CFBE41o-human bronchial epithelial cell lines. 27 The cells were transfected with F508del-CFTR at 37 C, then incubated with cysteamine (250 mM) for 18 h, which was washed out, followed by 24 to 48 h of culture, according to a previously described procedure.…”

Section: Resultsmentioning

confidence: 99%

“…We discovered that cystamine, a small molecule that acts as a modulator of proteostasis, is able to rescue and stabilize F508del-CFTR at the PM of CFBE41o-airway epithelial cell lines as well as in primary human nasal epithelial cells. 27,28,31 In this study we evaluated the therapeutic potential of cysteamine, the reduced form of cystamine, stimulated by the fact that cysteamine is an FDAapproved drug with a known safety profile and good bioavailability upon oral administration. 57 In a preclinical animal model, the Cftr F508del mouse, we observed that the oral treatment for 5 wk with cysteamine significantly reduced mortality, improved weight gain, and increased the expression of functional CFTR protein at the intestinal level, at the same time that it restored BECN1 protein expression to wild-type levels.…”

Section: Discussionmentioning

confidence: 99%

“…[26][27][28] Dysfunctional F508del-CFTR protein induces persistent activation of TGM2 (transglutaminase 2), 29,30 resulting in cross-linking of several TGM2 substrates, including BECN1, 26 a protein essential for autophagy. This leads to functional sequestration of the BECN1 interactome in intracellular aggregates, resulting in defective autophagy with consequent accumulation of the autophagic substrate SQSTM1.…”

Section: F508delmentioning

confidence: 99%

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Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

et al. 2014

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These authors equally contributed to this work.Keywords: cystic fibrosis, CFTR, autophagy, cysteamine, epigallocatechin gallate, sweat chloride Abbreviations: BECN1/Beclin 1, autophagy-related; CF, cystic fibrosis; CFTR, cystic fibrosis transmembrane conductance regulator; CHX, cycloheximide; CSNK2, casein kinase 2; CXCL2, chemokine (C-X-C motif) ligand 2; CXCL8, chemokine (C-X-C motif) ligand 8; EGCG, epigallocatechin gallate; FEV, forced expiratory volume; PM, plasma membrane; RPD, rectal potential difference; SQSTM1, sequestosome 1; TGM2, transglutaminase 2; TNF, tumor necrosis factor.Restoration of BECN1/Beclin 1-dependent autophagy and depletion of SQSTM1/p62 by genetic manipulation or autophagy-stimulatory proteostasis regulators, such as cystamine, have positive effects on mouse models of human cystic fibrosis (CF). These measures rescue the functional expression of the most frequent pathogenic CFTR mutant, F508del, at the respiratory epithelial surface and reduce lung inflammation in Cftr F508del homozygous mice. Cysteamine, the reduced form of cystamine, is an FDA-approved drug. Here, we report that oral treatment with cysteamine greatly reduces the mortality rate and improves the phenotype of newborn mice bearing the F508del-CFTR mutation. Cysteamine was also able to increase the plasma membrane expression of the F508del-CFTR protein in nasal epithelial cells from F508del homozygous CF patients, and these effects persisted for 24 h after cysteamine withdrawal. Importantly, this cysteamine effect after washout was further sustained by the sequential administration of epigallocatechin gallate (EGCG), a green tea flavonoid, both in vivo, in mice, and in vitro, in primary epithelial cells from CF patients. In a pilot clinical trial involving 10 F508del-CFTR homozygous CF patients, the combination of cysteamine and EGCG restored BECN1, reduced SQSTM1 levels and improved CFTR function from nasal epithelial cells in vivo, correlating with a decrease of chloride concentrations in sweat, as well as with a reduction of the abundance of TNF/ TNF-alpha (tumor necrosis factor) and CXCL8 (chemokine [C-X-C motif] ligand 8) transcripts in nasal brushing and TNF and CXCL8 protein levels in the sputum. Altogether, these results suggest that optimal schedules of cysteamine plus EGCG might be used for the treatment of CF caused by the F508del-CFTR mutation.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: F508delmentioning

confidence: 99%

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Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

et al. 2014

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“…It is well known that pancreatic β-cells undergo extensive proinsulin biosynthesis, and, therefore, face a high protein-folding burden. In such conditions, pancreatic β-cell survival is highly dependent on the unfolded protein response machinery that is compromised in CF patients due to the accumulation of misfolded CFTR protein and/or failure of autophagy [51][52][53] . Recent studies have revealed that etiologic CFTR mutations trigger a cascade of events that culminate in protein Beclin 1 depletion and autophagy impairment [51,52] .…”

Section: Discussionmentioning

confidence: 99%

Steady-State Therapy with Azithromycin or Low-Dose Prednisolone in Paediatric Cystic Fibrosis Patients: Inflammatory Markers and Disease Progression

Shmarina

Pukhalsky²,

Avakian³

et al. 2017

Int Arch Allergy Immunol

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Background: Anti-inflammatory therapy is a logical approach to slowing the inevitable lung function deterioration in cystic fibrosis (CF) patients. This study's aim was to evaluate inflammatory markers and disease progression in paediatric CF patients chronically treated with azithromycin or low-dose prednisolone. Methods: The study included 204 patients with CF and 100 healthy controls; 102 CF patients were treated with basic therapy only (without anti-inflammatory treatment; WAT), and 102 individuals received basic therapy along with azithromycin (n = 59) or low-dose prednisolone (n = 43). The median duration of therapy was 24 months (range 12-82) with azithromycin and 31 months (range 12-180) with prednisolone. A cross-sectional analysis of plasma and sputum biomarkers was performed. Results: Compared with the healthy controls, the WAT group showed elevated IFN-γ, IL-10 (total), and TGFβ₁ concentrations, and decreased TNFα (total) and adrenocorticotropic hormone (ACTH) levels (all p < 0.05). Plasma TNFα (total) concentrations in azithromycin/prednisolone patients were significantly higher than those in WAT patients and similar to those of healthy children. In contrast, IL-10 (total) levels were significantly decreased in azithromycin/prednisolone-treated patients compared with WAT patients. Children from the azithromycin group demonstrated ACTH levels similar to those of healthy controls. Azithromycin-treated patients showed a significantly reduced rate of CF-related liver disease and a significantly increased incidence of glucose metabolism disturbances. Conclusions: Steady-state anti-inflammatory treatments may have a sustained immunomodulatory action at systemic and local levels in CF patients. Further investigations are needed to assess the effects of supportive azithromycin therapy on the hypothalamic-pituitary-adrenal axis and the incidence of non-pulmonary CF complications.

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“…[23] Similarly, difference in its susceptibility and kinetics to inhibitors under various experimental conditions has also been observed. [22] Therefore, it becomes interesting in studying polyol pathway in different tissue sites under various influencing conditions to identify variations in activity profile of inhibitors.…”

Section: Resultsmentioning

confidence: 98%

Vegetables′ juice influences polyol pathway by multiple mechanisms in favour of reducing development of oxidative stress and resultant diabetic complications

et al. 2014

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Objective:Hyperglycemia induced generation of free radicals and consequent development of oxidative stress by polyol pathway is one of the crucial mechanisms stirring up development of diabetic complications. We evaluated influence of ten vegetables’ juice on polyol pathway along with their antioxidant and antioxidative stress potentials.Materials and Methods:Aldose reductase activity was determined utilising goat lens and human erythrocytes. In goat lens, utilization of nicotinamine adenine dinucleotide phosphate (NADPH) and aldose reductase inhibition was assayed. In human erythrocytes, sorbitol formation was measured as an index of aldose reductase activity under normoglycemic and hyperglycemic conditions. Ability of juices in inhibiting oxidative damage to deoxyribose sugar and calf thymus DNA and inhibitory activity against hydrogen peroxide induced hemolysis of erythrocytes was also analysed. Phytochemical contents like total polyphenol, total flavonoid and total protein were measured to find their influence on biological activities.Results:Vegetables’ juice displayed varying degrees of inhibitory potentials in mitigating NADPH dependent catalytic activity of aldose reductase in goat lens, accumulation of sorbitol in human erythrocytes under different glucose concentrations; Fenton-reaction induced oxidative damage to deoxyribose sugar, and calf thymus DNA. Substantial variations in vegetables phytochemicals content were also noticed in this study.Conclusions:Vegetables’ juice possesses potent activities in influencing polyol pathway by various mechanisms in favour of reducing development of oxidative stress independent of their inherent antioxidative properties. Juice of ivy gourd followed by green cucumber and ridge gourd were among the most potent for they displayed strong activities on various parameters analysed in this study. These vegetables’ juice may become part of mechanism-based complementary antioxidant therapy to prevent development of diabetic complications.

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Targeting the Intracellular Environment in Cystic Fibrosis: Restoring Autophagy as a Novel Strategy to Circumvent the CFTR Defect

Cited by 116 publications

References 118 publications

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

Restoration of CFTR function in patients with cystic fibrosis carrying the F508del-CFTR mutation

Steady-State Therapy with Azithromycin or Low-Dose Prednisolone in Paediatric Cystic Fibrosis Patients: Inflammatory Markers and Disease Progression

Vegetables′ juice influences polyol pathway by multiple mechanisms in favour of reducing development of oxidative stress and resultant diabetic complications

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