The online version of this article (https://doi.org/10.1186/s12931-017-0705-8) contains supplementary material, which is available to authorized users.
Chronic lung disease resulting from dysfunctional cystic fibrosis transmembrane conductance regulator (CFTR) and NFκB-mediated neutrophilic-inflammation forms the basis of CF-related mortality. Here we aimed to evaluate if HDAC inhibition controls Pseudomonas-aeruginosa-lipopolysaccharide (Pa-LPS) induced airway inflammation and CF-lung disease.
For in vitro experiments, HEK293-cells were transfected with IL-8 or NFκB-firefly luciferase, and SV40-renilla- luciferase reporter constructs or ΔF508-CFTR-pCEP, followed by treatment with suberoylanilide hydroxamic acid (SAHA), Trichostatin-A (TSA) and/or TNFα. For murine studies, Cftr +/+ or Cftr −/− mice (n = 3) were injected/instilled with Pa-LPS and/or treated with SAHA or vehicle control. The progression of lung disease was monitored by quantifying changes in inflammatory markers (NFκB), cytokines (IL-6/IL-10), neutrophil activity (MPO, myeloperoxidase and/or NIMP-R14) and T-reg numbers.
SAHA treatment significantly (p < 0.05) suppresses TNFα-induced NFκB and IL-8 reporter activities in HEK293-cells. Moreover, SAHA, Tubacin (selective HDAC6-inhibitor) or HDAC6-shRNAs controls CSE-induced ER-stress activities (p < 0.05). In addition, SAHA restores trafficking of misfolded-ΔF508-CFTR, by inducing protein levels of both B and C forms of CFTR. Murine studies using Cftr +/+ or Cftr −/− mice verified that SAHA controls Pa-LPS induced IL-6 levels, and neutrophil (MPO levels and/or NIMP-R14), NFκB-(inflammation) and Nrf2 (oxidative-stress marker) activities, while promoting FoxP3+ T-reg activity.
In summary, SAHA-mediated HDAC inhibition modulates innate and adaptive immune responses involved in pathogenesis and progression of inflammatory CF-lung disease.
Additional file 1: Figure S1. Selective HDAC inhibition controls ER-stress activity. (A) HEK293 cells were transiently transfected with two different HDAC6 shRNA constructs or control plasmid and the cell lysates were immunoblotted for HDAC6 and β-actin. The immunoblot verifies the >2 fold knockdown efficiency of HDACshRNA for experiment shown in B. (B) HEK293 cells were transiently transfected with a secretory gaussia reporter plasmid and/or HDAC6 shRNA. After 6 h of transfection cells were treated with CSE (cigarette smoke extract, as ER stress activator), SAHA (10 μM), or Tubacin (10 μM) after 6 h. At 72 (B, upper panel) or 24 (B, lower panel) hours, supernatants were collected and read with the aforementioned Dual Luciferase Reporter System to determine ER-stress activity. The data (mean ± SD of triplicate samples) shows that CSE induced ER-stress activity (p < 0.05) is significantly controlled by Class II HDAC inhibitor, (SAHA), selective HDAC6 inhibitor (Tubacin, B-upper panel) and HDAC6 shRNA (B-lower panel) suggesting the therapeutic potential of selective HDACi in controlling CF-related ER-stress response. (C) The Cftr +/+ mice were exposed to sub-chronic cigarette smoke (sc-CS, 8 weeks) and i.t. instilled with SAHA (50 μg/mouse, three total doses with one day interval before the termination of the experiment). The BALF cells were harvested and analyzed for CD4+ (CD4-PE antibody) and FoxP3+ [(FoxP3 (Rabbit polyclonal primary Ab)-Anti-Rabbit-FITC (secondary Ab)] cells by flow cytometry using the BD FACS Caliber instrument. The data indicates that SAHA treatment induces the levels of FoxP3+ T regs to counteract the sc-CS mediated airway inflammation. (JPEG 259 kb)12931_2017_705_MOESM1_ESM.jpg
Hutt DM, Olsen CA, Vickers CJ, Herman D, Chalfant M, Montero A, Leman LJ, Burkle R, Maryanoff BE, Balch WE, Ghadiri MR. Potential agents for treating cystic fibrosis: cyclic Tetrapeptides that restore trafficking and activity of DeltaF508-CFTR. ACS Med Chem Lett. 2011;2:703–7. CrossRefPubMedPubMedCentral
Sosnay PR, Raraigh KS, Gibson RL. Molecular genetics of cystic fibrosis Transmembrane conductance regulator: genotype and phenotype. Pediatr Clin N Am. 2016;63:585–98. CrossRef
Luciani A, Villella VR, Esposito S, Brunetti-Pierri N, Medina D, Settembre C, Gavina M, Pulze L, Giardino I, Pettoello-Mantovani M, et al. Defective CFTR induces aggresome formation and lung inflammation in cystic fibrosis through ROS-mediated autophagy inhibition. Nat Cell Biol. 2010;12:863–75. CrossRefPubMed
Vij N, Min T, Bodas M, Gorde A, Roy I. Neutrophil targeted nano-drug delivery system for chronic obstructive lung diseases. Nanomedicine. 2016; In Press
Iannitti RG, Carvalho A, Cunha C, De Luca A, Giovannini G, Casagrande A, Zelante T, Vacca C, Fallarino F, Puccetti P, et al. Th17/Treg imbalance in murine cystic fibrosis is linked to indoleamine 2,3-dioxygenase deficiency but corrected by kynurenines. Am J Respir Crit Care Med. 2013;187:609–20. CrossRefPubMed
Min T, Bodas M, Mazur S, Vij N. Critical role of proteostasis-imbalance in pathogenesis of COPD and severe emphysema. J Mol Med (Berl). 2011;89:577–93. CrossRef
Dinarello CA. Inhibitors of histone deacetylases as anti-inflammatory drugs. Ernst Schering Res Found Workshop. 2006:45–60.
Anil N, Singh M. CD4(+)CD25(high) FOXP3(+) regulatory T cells correlate with FEV1 in north Indian children with cystic fibrosis. Immunol Investig. 2014;43:535–43. CrossRef
Stanton BA. Adverse effects of pseudomonas aeruginosa on CFTR chloride secretion and the host immune response. Am J Physiol Cell Physiol. 2017;312(4):C357-C366. https://doi.org/10.1152/ajpcell.00373.2016. Epub 2017 Jan 25
Stanton BA. Adverse effects of Pseudomonas Aeruginosa on CFTR chloride secretion and the host immune response. Am J Physiol Cell Physiol. 2017; ajpcell 00373 02016
Cebotaru L, Vij N, Ciobanu I, Wright J, Flotte T, Guggino WB. Cystic fibrosis transmembrane regulator missing the first four transmembrane segments increases wild type and DeltaF508 processing. J Biol Chem. 2008;283(32):21926-33. https://doi.org/10.1074/jbc.M709156200. Epub 2008 May 28.
Vij N. Nano-based rescue of dysfunctional autophagy in chronic obstructive lung diseases. Expert Opin Drug Deliv. 2016:1–7.
- Inhibition of histone-deacetylase activity rescues inflammatory cystic fibrosis lung disease by modulating innate and adaptive immune responses
- BioMed Central
Neu im Fachgebiet Innere Medizin
Meistgelesene Bücher aus der Inneren Medizin
Mail Icon II