Cystic fibrosis (CF) is the most common genetic disease in Caucasians. CF is manifested by abnormal accumulation of mucus in the lungs, which serves as fertile ground for the growth of microorganisms leading to recurrent infections and ultimately, lung failure. Mucus in CF patients consists of DNA from dead neutrophils as well as mucins produced by goblet cells. MUC5AC mucin leads to pathological plugging of the airways whereas MUC5B has a protective role against bacterial infection. Therefore, decreasing the level of MUC5AC while maintaining MUC5B intact would in principle be a desirable mucoregulatory treatment outcome. Fenretinide prevented the lipopolysaccharide-induced increase of MUC5AC gene expression, without affecting the level of MUC5B, in a lung goblet cell line. Additionally, fenretinide treatment reversed the pro-inflammatory imbalance of fatty acids by increasing docosahexaenoic acid and decreasing the levels of arachidonic acid in a lung epithelial cell line and primary leukocytes derived from CF patients. Furthermore, for the first time we also demonstrate the effect of fenretinide on multiple unsaturated fatty acids, as well as differential effects on the levels of long- compared to very-long-chain saturated fatty acids which are important substrates of complex phospholipids. Finally, we demonstrate that pre-treating mice with fenretinide in a chronic model of P. aeruginosa lung infection efficiently decreases the accumulation of mucus. These findings suggest that fenretinide may offer a new approach to therapeutic modulation of pathological mucus production in CF.
- MeSH
- aplikace orální MeSH
- buněčné linie MeSH
- cystická fibróza komplikace genetika patologie MeSH
- fenretinid aplikace a dávkování MeSH
- fosfolipidy metabolismus MeSH
- hlen metabolismus MeSH
- krysa rodu rattus MeSH
- kyselina arachidonová metabolismus MeSH
- kyseliny dokosahexaenové metabolismus MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- mucin 5AC metabolismus MeSH
- mucin 5B metabolismus MeSH
- myši inbrední CFTR MeSH
- myši MeSH
- plíce účinky léků metabolismus patologie MeSH
- pneumonie mikrobiologie patologie prevence a kontrola MeSH
- pseudomonádové infekce mikrobiologie patologie prevence a kontrola MeSH
- Pseudomonas aeruginosa patogenita MeSH
- respirační sliznice cytologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cystic fibrosis (CF) is the most common autosomal-recessive disease in Caucasians caused by mutations in the CF transmembrane regulator (CFTR) gene. Patients are usually diagnosed in infancy and are burdened with extensive medical treatments throughout their lives. One of the first documented biochemical defects in CF, which predates the cloning of CFTR gene for almost three decades, is an imbalance in the levels of polyunsaturated fatty acids (PUFAs). The principal hallmarks of this imbalance are increased levels of arachidonic acid and decreased levels of docosahexaenoic acids (DHA) in CF. This pro-inflammatory profile of PUFAs is an important component of sterile inflammation in CF, which is known to be detrimental, rather than protective for the patients. Despite decades of intensive research, the mechanistic basis of this phenomenon remains unclear. In this review we summarized the current knowledge on the biochemistry of PUFAs, with a focus on the metabolism of AA and DHA in CF. Finally, a synthetic retinoid called fenretinide (N-(4-hydroxy-phenyl) retinamide) was shown to be able to correct the pro-inflammatory imbalance of PUFAs in CF. Therefore, its pharmacological actions and clinical potential are briefly discussed as well.
- MeSH
- antiflogistika farmakologie terapeutické užití MeSH
- cystická fibróza farmakoterapie metabolismus MeSH
- esenciální mastné kyseliny metabolismus MeSH
- fenretinid farmakologie terapeutické užití MeSH
- lidé MeSH
- nenasycené mastné kyseliny metabolismus MeSH
- zánět farmakoterapie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Zona pellucida binding protein 2 (Zpbp2) and ORMDL sphingolipid biosynthesis regulator 3 (Ormdl3), mapped downstream of Zpbp2, were identified as two genes associated with airway hyper-responsiveness (AHR). Ormdl3 gene product has been shown to regulate the biosynthesis of ceramides. Allergic asthma was shown to be associated with an imbalance between very-long-chain ceramides (VLCCs) and long-chain ceramides (LCCs). We hypothesized that Fenretinide can prevent the allergic asthma-induced augmentation of Ormdl3 gene expression, normalize aberrant levels of VLCCs and LCCs, and treat allergic asthma symptoms. We induced allergic asthma by house dust mite (HDM) in A/J WT mice and Zpbp2 KO mice expressing lower levels of Ormdl3 mRNA than WT. We investigated the effect of a novel formulation of Fenretinide, LAU-7b, on the AHR, inflammatory cell infiltration, mucus production, IgE levels, and ceramide levels. Although lower Ormdl3 expression, which was observed in Zpbp2 KO mice, was associated with lower AHR, allergic Zpbp2 KO mice were not protected from inflammatory cell infiltration, mucus accumulation, or aberrant levels of VLCCs and LCCs induced by HDM. LAU-7b treatment protects both the Zpbp2 KO and WT mice. The treatment significantly lowers the gene expression of Ormdl3, normalizes the VLCCs and LCCs, and corrects all the other phenotypes associated with allergic asthma after HDM challenge, except the elevated levels of IgE. LAU-7b treatment prevents the augmentation of Ormdl3 expression and ceramide imbalance induced by HDM challenge and protects both WT and Zpbp2 KO mice against allergic asthma symptoms. SIGNIFICANCE STATEMENT: Compared with A/J WT mice, KO mice with Zpbp2 gene deletion have lower AHR and lower levels of Ormdl3 expression. The novel oral clinical formulation of Fenretinide (LAU-7b) effectively lowers the AHR and protects against inflammatory cell infiltration and mucus accumulation induced by house dust mite in both Zpbp2 KO and WT A/J mice. LAU-7b prevents Ormdl3 overexpression in WT allergic mice and corrects the aberrant levels of very-long-chain and long-chain ceramides in both WT and Zpbp2 KO allergic mice.
- MeSH
- bronchiální astma farmakoterapie metabolismus MeSH
- ceramidy metabolismus MeSH
- down regulace účinky léků MeSH
- exprese genu účinky léků MeSH
- fenretinid farmakologie MeSH
- membránové proteiny metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši knockoutované MeSH
- myši MeSH
- respirační alergie farmakoterapie metabolismus MeSH
- zánět metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
PURPOSE: To assess the efficacy of the novel clinical formulation of fenretinide (LAU-7b) for the treatment of allergic asthma. To study the association between LAU-7b treatment in allergic asthma and the modulation of very long chain ceramides (VLCC). METHODS: We used two allergens (OVA and HDM) to induce asthma in mouse models and we established a treatment protocol with LAU-7b. The severity of allergic asthma reaction was quantified by measuring the airway resistance, quantifying lung inflammatory cell infiltration (Haematoxylin and eosin stain) and mucus production (Periodic acid Schiff satin). IgE levels were measured by ELISA. Immunophenotyping of T cells was done using Fluorescence-activated cell sorting (FACS) analysis. The analysis of the specific species of lipids and markers of oxidation was performed using mass spectrometry. RESULTS: Our data demonstrate that 10 mg/kg of LAU-7b was able to protect OVA- and HDM-challenged mice against increase in airway hyperresponsiveness, influx of inflammatory cells into the airways, and mucus production without affecting IgE levels. Treatment with LAU-7b significantly increased percentage of regulatory T cells and CD4+ IL-10-producing T cells and significantly decreased percentage of CD4+ IL-4-producing T cells. Our data also demonstrate a strong association between the improvement in the lung physiology and histology parameters and the drug-induced normalization of the aberrant distribution of ceramides in allergic mice. CONCLUSION: 9 days of 10 mg/kg of LAU-7b daily treatment protects the mice against allergen-induced asthma and restores VLCC levels in the lungs and plasma.
- MeSH
- alergeny imunologie MeSH
- bronchiální astma farmakoterapie imunologie metabolismus MeSH
- ceramidy metabolismus MeSH
- fenretinid terapeutické užití MeSH
- klinické protokoly MeSH
- methylcelulosa chemie MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- ovalbumin imunologie MeSH
- příprava léků MeSH
- Pyroglyphidae imunologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
This project is aimed at a detailed analysis of the expression of five candidate biomarkers (PBX1, HOXC9, NF1, HMGI-C and HMGA2) associated with the resistance to retinoids in neuroblastoma cells. Endogenous expression of these biomarkers should be evaluated in established neuroblastoma cell lines as well as in paired samples of tumor tissue taken from the individual patients using PCR and immunodetection methods. At least 25-30 paired tumor samples and 25 neuroblastoma cell lines should be included in this study. Possible differences in expression of biomarkers between paired samples taken from the same patient at diagnosis and after induction chemotherapy should be verified on cell lines treated with selected retinoids. This study brings especially completely new information based on multiple analysis of all candidate biomarkers known to be associated with the resistance to retinoids in one experimental cohort. Obtained data may help to identify useful predictive biomarkers of the patient’s response to retinoids within high-risk neuroblastoma category.
Projekt je zaměřen na detailní analýzu exprese pěti kandidátních biomarkerů (PBX1, HOXC9, NF1, HMGI-C a HMGA2), které souvisejí s rezistencí k retinoidům v buňkách neuroblastomu. Endogenní exprese těchto biomarkerů bude hodnocena pomocí PCR a imunodetekce ve stabilizovaných neuroblastomových buněčných liniích a v párových vzorcích nádorové tkáně od jednotlivých pacientů. Do studie bude zařazeno nejméně 25-30 párových vzorků nádorové tkáně odebírané pacientům a 25 neuroblastomových buněčných linií. Případné rozdíly v expresi biomarkerů mezi párovými vzorky téhož pacienta, které byly odebrány v době diagnózy a po indukční chemoterapii, budou ověřeny pomocí ovlivnění buněčných linií vybranými retinoidy. Tato studie přinese zejména zcela nové poznatky založené na komplexní analýze všech kandidátních biomarkerů rezistence k retinoidům v rámci jedné experimentální kohorty. Získané informace tak mohou přispět k identifikaci prediktivních biomarkerů, které budou v praxi využitelné ke stanovení léčebné odpovědi na retinoidy u pacientů s vysoce rizikovými neuroblastomy.
- MeSH
- bexaroten MeSH
- chemorezistence MeSH
- fenretinid MeSH
- imunologické techniky MeSH
- nádorové biomarkery MeSH
- neuroblastom MeSH
- neurofibromin 1 chemie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- pre-B-buněčný leukemický transkripční faktor 1 chemie MeSH
- proteiny HMGA chemie MeSH
- retinoidy MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- onkologie
- pediatrie
- chemie, klinická chemie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
Although the administration of retinoids represents an important part of treatment for children suffering from high-risk neuroblastomas, approximately 50% of these patients do not respond to this therapy or develop resistance to retinoids during treatment. Our study focused on the comparative analysis of the expression of five genes and corresponding proteins (DDX39A, HMGA1, HMGA2, HOXC9 and PBX1) that have recently been discussed as possible predictive biomarkers of clinical response to retinoid differentiation therapy. Expression of these five candidate biomarkers was evaluated at both the mRNA and protein level in the same subset of 8 neuroblastoma cell lines after treatment with natural or synthetic retinoids. We found that the cell lines that were HMGA2-positive and/or HOXC9-negative have a reduced sensitivity to retinoids. Furthermore, the experiments revealed that the retinoid-sensitive cell lines showed a uniform pattern of change after treatment with both natural and sensitive retinoids: increased DDX39A and decreased PBX1 protein levels. Our results showed that in NBL cells, these putative protein biomarkers are associated with sensitivity or resistance to retinoids, and their endogenous or induced expression can distinguish between these two phenotypes.
- MeSH
- antitumorózní látky farmakologie MeSH
- bexaroten farmakologie MeSH
- biomarkery farmakologické metabolismus MeSH
- chemorezistence účinky léků genetika MeSH
- DEAD-box RNA-helikasy genetika metabolismus MeSH
- dítě MeSH
- fenretinid farmakologie MeSH
- fixace tkání MeSH
- homeodoménové proteiny genetika metabolismus MeSH
- isotretinoin farmakologie MeSH
- kojenec MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- nádorové buněčné linie MeSH
- nádory nervového systému genetika metabolismus patologie chirurgie MeSH
- neuroblastom genetika metabolismus patologie chirurgie MeSH
- novorozenec MeSH
- pre-B-buněčný leukemický transkripční faktor 1 genetika metabolismus MeSH
- předškolní dítě MeSH
- proliferace buněk účinky léků MeSH
- protein HMGA1A genetika metabolismus MeSH
- protein HMGA2 genetika metabolismus MeSH
- tretinoin analogy a deriváty farmakologie MeSH
- zalévání tkání do parafínu MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cystic fibrosis is the most common genetic disease, in which symptoms may be alleviated but not fully eliminated. Ceramides have long been implicated in the inflammatory etiology of cystic fibrosis, with contradicting reports with regards to their role. Recently, significant biological and biophysical differences have been observed between long- and very long-chain ceramides. This work reveals that long-chain ceramides are upregulated whereas very long-chain ceramides are downregulated in cell lines, mouse animal model, and patients with cystic fibrosis, compared with their controls. Treatment with fenretinide decreases the levels of long-chain ceramides and increases the levels of very long-chain ceramides. Our results show that restoration of cystic fibrosis conductance regulator (CFTR) expression is associated with normalization of aberrant levels of specific ceramides. This demonstrates for the first time a correlation between CFTR protein expression and regulation of specific ceramide levels. Furthermore, using cystic fibrosis lung epithelial cell lines, we demonstrate that this effect can be attributed to the transcriptional downregulation of ceramide synthase 5 (Cers5) enzyme. We also discovered a partial synergism between fenretinide and zinc (Zn2+), which deficiency has been reported in patients with cystic fibrosis. Overall, in addition to having direct translational application, we believe that our findings contribute to the understanding of ceramide metabolism in cystic fibrosis, as well as other inflammatory diseases where imbalances of ceramides have also been observed. KEY MESSAGES: Long- and very long-chain ceramides (LCCs and VLCCs) are biochemically distinct. LCCs are upregulated whereas VLCCs are downregulated in cystic fibrosis. Fenretinide downregulates the levels of LCCs and upregulates the levels of VLCCs. Fenretinide changes the balance of LCCs and VLCCs by downregulating Cers5 enzyme. Fenretinide and zinc ions cooperate in the modulation of ceramide levels.
- MeSH
- aktivace transkripce účinky léků MeSH
- buněčné linie MeSH
- ceramidy analýza krev metabolismus MeSH
- cystická fibróza krev farmakoterapie metabolismus MeSH
- dospělí MeSH
- down regulace účinky léků MeSH
- fenretinid terapeutické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- PPAR gama agonisté MeSH
- sfingosin-N-acyltransferasa antagonisté a inhibitory genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- MeSH
- chemie farmaceutická * metody MeSH
- disulfiram terapeutické užití MeSH
- fenretinid terapeutické užití MeSH
- kyselina valproová terapeutické užití MeSH
- lidé MeSH
- metformin terapeutické užití MeSH
- nádory * farmakoterapie MeSH
- přehodnocení terapeutických indikací léčivého přípravku * MeSH
- thalidomid terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- androstadieny terapeutické užití MeSH
- antiflogistika nesteroidní terapeutické užití MeSH
- antikarcinogenní látky terapeutické užití MeSH
- antitumorózní látky hormonální škodlivé účinky terapeutické užití MeSH
- bisfosfonáty terapeutické užití MeSH
- fenretinid terapeutické užití MeSH
- inhibitory aromatasy škodlivé účinky terapeutické užití MeSH
- konsensuální konference jako téma MeSH
- lidé MeSH
- metformin terapeutické užití MeSH
- modulátory estrogenních receptorů škodlivé účinky terapeutické užití MeSH
- nádory prsu prevence a kontrola MeSH
- nitrily terapeutické užití MeSH
- norpregneny terapeutické užití MeSH
- piperidiny terapeutické užití MeSH
- premenopauza MeSH
- pyrrolidiny terapeutické užití MeSH
- raloxifen hydrochlorid terapeutické užití MeSH
- retinoidy terapeutické užití MeSH
- selektivní modulátory estrogenních receptorů škodlivé účinky terapeutické užití MeSH
- statiny terapeutické užití MeSH
- tamoxifen terapeutické užití MeSH
- tetrahydronaftaleny terapeutické užití MeSH
- thiofeny terapeutické užití MeSH
- triazoly MeSH
- znalecký posudek MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH