Cigarette smoke exposure is associated with vitamin D3 deficiencies in patients with chronic rhinosinusitis.

BACKGROUND: Cigarette smoke (CS) plays a role in the exacerbation of chronic rhinosinusitis (CRS); however, the mechanism for this is unknown. We hypothesize that CS impairs human sinonasal epithelial cell (HSNEC) conversion of 25(OH)D3 (25VD3) to 1,25-dihydroxyvitamin D3 (1,25VD3) and, furthermore, that supplementation with 1,25VD3 will reverse smoke-induced inflammatory responses by HSNECs. OBJECTIVE: We sought to determine the effect of CS on vitamin D3 (VD3) levels, conversion, and regulation of CS-induced inflammation in control subjects and patients with CRS. METHODS: Blood and sinus tissue explants were collected at the time of surgery from control subjects, patients with chronic rhinosinusitis without nasal polyps, and patients with chronic sinusitis with nasal polyps (CRSwNP). Expression of VD3 metabolizing enzymes were measured by using RT-PCR. Primary HSNECs were cultured from tissue explants. 25VD3 with and without cigarette smoke extract (CSE) was used to examine conversion of 25VD3 to 1,25VD3, as well as HSNEC production of proinflammatory cytokines. RESULTS: CS exposure was associated with reduced circulating and sinonasal 25VD3 levels in all groups compared with those seen in CS-naive, disease-matched counterparts. CS exposure decreased expression of CYP27B1 and was especially pronounced in patients with CRSwNP. CSE impairs control HSNEC conversion of 25VD3. HSNECs from patients with CRSwNP also demonstrate an intrinsic reduction in conversion of 25VD3 to 1,25VD3. Exogenous 1,25VD3 reduces CSE-induced cytokine production by HSNECs. CONCLUSIONS: Exposure to CS is associated with reduced 25VD3 levels and an impaired ability of HSNECs to convert 25VD3 to 1,25VD3. Addition of 1,25VD3 reduces the proinflammatory effects of CS on HSNECs. Impaired VD3 conversion by CS exposure represents a novel mechanism through which CS induces its proinflammatory effects.

Systemic monocyte-derived dendritic cells and associated Th2 skewing in chronic rhinosinusitis.

INTRODUCTION: Monocyte-derived dendritic cells (moDCs) are antigen-presenting cells capable of directing immune responses toward T-helper 1 (Th1) or T-helper 2 (Th2) phenotypes. The systemic profile of moDCs and their association with Th1/Th2 skewing in chronic rhinosinusitis (CRS) is unclear. The purpose of this study is to characterize circulating moDCs in controls, CRS without nasal polyps (CRSsNP), and CRS with nasal polyps (CRSwNP) and correlate moDCs with Th1/Th2 skewing, mucosal inflammation on computed tomography (CT), and quality of life (QoL). STUDY DESIGN: Cross-sectional study. SETTING: Tertiary care hospital. SUBJECTS: Blood was drawn from control (n = 12), CRSsNP (n = 18), and CRSwNP (n = 15) patients during endoscopic sinus surgery. METHODS: Peripheral blood moDCs were analyzed with flow cytometry for expression of HLA-DR, CD209, and CD14. Th1 and Th2 cells were identified by CXCR3 and CCR8 expression, respectively. Lund-Mackay CT scores were assigned by blinded graders. Sino-Nasal Outcome Test 22 (SNOT-22) surveys were completed by patients before surgery. RESULTS: CRSsNP and CRSwNP displayed elevations in systemic moDCs compared with controls. In CRSwNP, systemic Th2 skewing was observed and circulating CD4+ Th2 cells correlated with percent moDCs. MoDCs strongly correlated with higher Lund-Mackay CT scores in CRSsNP but not in CRSwNP. No relationship between moDCs and SNOT-22 scores was observed for either subset of CRS. CONCLUSION: These data support that CRSwNP and CRSsNP display alterations in systemic immune profiles. CRSwNP is characterized by significant elevations in circulating moDCs, which is associated with systemic Th2-biased inflammation. Circulating moDCs are associated with mucosal inflammation on CT imaging in CRSsNP. No association between moDCs and QoL is evident in either CRS subset.

Phosphoproteomic profiling of human myocardial tissues distinguishes ischemic from non-ischemic end stage heart failure.

The molecular differences between ischemic (IF) and non-ischemic (NIF) heart failure are poorly defined. A better understanding of the molecular differences between these two heart failure etiologies may lead to the development of more effective heart failure therapeutics. In this study extensive proteomic and phosphoproteomic profiles of myocardial tissue from patients diagnosed with IF or NIF were assembled and compared. Proteins extracted from left ventricular sections were proteolyzed and phosphopeptides were enriched using titanium dioxide resin. Gel- and label-free nanoscale capillary liquid chromatography coupled to high resolution accuracy mass tandem mass spectrometry allowed for the quantification of 4,436 peptides (corresponding to 450 proteins) and 823 phosphopeptides (corresponding to 400 proteins) from the unenriched and phospho-enriched fractions, respectively. Protein abundance did not distinguish NIF from IF. In contrast, 37 peptides (corresponding to 26 proteins) exhibited a ≥ 2-fold alteration in phosphorylation state (p<0.05) when comparing IF and NIF. The degree of protein phosphorylation at these 37 sites was specifically dependent upon the heart failure etiology examined. Proteins exhibiting phosphorylation alterations were grouped into functional categories: transcriptional activation/RNA processing; cytoskeleton structure/function; molecular chaperones; cell adhesion/signaling; apoptosis; and energetic/metabolism. Phosphoproteomic analysis demonstrated profound post-translational differences in proteins that are involved in multiple cellular processes between different heart failure phenotypes. Understanding the roles these phosphorylation alterations play in the development of NIF and IF has the potential to generate etiology-specific heart failure therapeutics, which could be more effective than current therapeutics in addressing the growing concern of heart failure.

Differential expression of adhesion molecules by sinonasal fibroblasts among control and chronic rhinosinusitis patients.

BACKGROUND: Chronic rhinosinusitis (CRS) is characterized by inflammatory cell migration into sinus tissue with resultant inflammation fueled by a milieu of cytokines. Fibroblasts may contribute to inflammation through expression of leukocyte adhesion molecules such as vascular cell adhesion molecule (VCAM) and intercellular adhesion molecule (ICAM). VCAM attracts eosinophils and mast cells contributing to Th2 skewing, and ICAM attracts neutrophils and to a lesser degree, eosinophils, and contributes to mixed Th1/Th2 skewing. The purpose of this study was to compare sinus fibroblast adhesion molecule expression ex vivo among CRS subtypes and in vitro after cytokine stimulation. METHODS: Sinus biopsy specimens were taken from control patients (n = 13), CRS without nasal polyposis (CRSsNP, n = 6), and CRS with nasal polyposis (CRSwNP, n = 15). ex vivo levels of VCAM and ICAM were measured by flow cytometry from single cell suspensions of tissue biopsy specimens. Changes in VCAM and ICAM expression to cytokine exposure were assessed using in vitro cultured sinonasal fibroblasts treated with tumor necrosis factor (TNF)-α, interleukin (IL)-4, or interferon (IFN)-γ. RESULTS: ex vivo VCAM expression was lowest in controls, higher in CRSsNP, and highest in CRSwNP. in vitro stimulation with TNF-α and IL-4, but not IFN-γ, increased VCAM among CRSsNP, while expression in CRSwNP remained elevated with all treatments except IFN-γ. ex vivo ICAM expression was elevated in both CRS subtypes. in vitro stimulation with TNF-α and IFN-γ, but not IL-4, increased ICAM expression in all patients with the largest effects among the CRSsNP subgroup. CONCLUSION: Sinonasal fibroblast expression of adhesion molecules in sinusitis varies by disease state and is selectively influenced by exposure to inflammatory cytokines.