IL-1ß induction of MUC5AC gene expression is mediated by CREB and NF-κB and repressed by dexamethasone.

Chronic airway diseases are characterized by inflammation and mucus overproduction. The MUC5AC mucin gene is upregulated by the proinflammatory cytokine interleukin-1 ß (IL-1ß) via activation of cAMP response element-binding protein (CREB) in the NCI-H292 cancer cell line and nuclear factor-κB (NF-κB) in the HBE1 transformed cell line, with each transcription factor binding to a cognate cis site in the proximal or distal region, respectively, of the MUC5AC promoter. We utilized primary differentiated human bronchial epithelial (HBE) and A549 lung adenocarcinoma cells to further investigate the contributions of CREB and NF-κB subunits to the IL-1ß-induced upregulation of MUC5AC. Data show that ligand binding of IL-1ß to the IL-1ß receptor is required to increase MUC5AC mRNA abundance. Chromatin immunoprecipitation analyses show direct binding of CREB to the previously identified cAMP response element site and binding of p65 and p50 subunits to a novel NF-κB site in a mucin-regulatory domain in the proximal promoter and to a previously identified NF-κB site in the distal promoter. P50 binds to both NF-κB sites at 1 h following IL-1ß exposure, but is replaced at 2 h by p65 in A549 cells and by a p50/p65 heterodimer in HBE cells. Thus IL-1ß activates multiple domains in the MUC5AC promoter but exhibits some cell-specific responses, highlighting the complexity of MUC5AC transcriptional regulation. Data show that dexamethasone, a glucocorticoid that transcriptionally represses MUC5AC gene expression under constitutive conditions, also represses IL-1ß-mediated upregulation of MUC5AC gene expression. A further understanding of mechanisms mediating MUC5AC regulation should lead to a honing of therapeutic approaches for the treatment of mucus overproduction in inflammatory lung diseases.

Increased poly(ADP-ribose) polymerase (PARP)-1 expression and activity are associated with inflammation but not goblet cell metaplasia in murine models of allergen-induced airway inflammation.

Inflammation plays a key role in lung injury and in the pathogenesis of asthma. Two murine models of allergic airway inflammation-sensitization and challenge to ovalbumin (OVA) and intratracheal exposure to interleukin-13 (IL13)-were used to evaluate the expression of poly(ADP-ribose) polymerase-1 (PARP-1) in allergic airway inflammation. Inflammation is prominent in OVA-induced allergic asthma, but this inflammation is greatly reduced by a PARP-1 inhibitor and almost eliminated when PARP-1 knockout mice are subjected to the OVA model. The present study temporally evaluated PARP-1 protein expression, localization, and activity, as well as inflammation and goblet cell metaplasia (GCM), in murine lungs following a single OVA challenge or IL13 exposure. Following OVA challenge PARP-1 protein expression and activity were greatly increased, being maximal at 3 to 5 days following OVA exposure and beginning to decrease by day 8. These changes correlated with the timing and degree of inflammation and GCM. In contrast, PARP-1 protein or activity did not change following single IL13 exposure, though GCM was manifested without inflammation. This study demonstrates that both PARP-1 protein and activity are increased by allergen-activated inflammatory mediators, excluding IL13, and that PARP-1 increase does not appear necessary for GCM, one of the characteristic markers of allergic airway inflammation in murine models.

Pharmacogenomics primer course for first professional year pharmacy students.

BACKGROUND: The importance of pharmacogenomics (PGx) is widely recognized in healthcare; however, PGx knowledge and confidence is lacking among many healthcare professionals. Pharmacists are the most logical PGx experts, yet most pharmacy schools do not require the basic science foundations of PGx as admission prerequisites. METHODS: A PGx primer course was developed for first year pharmacy students. In addition to genomics-based didactic material, this course also contains 'journaling' exercises and a group term paper based on a self-genotyping/phenotype exercise. The effectiveness of this course was evaluated by anonymous genomics knowledge surveys, course evaluations and unsolicited student comments. RESULTS: Data demonstrate this course engendered a significant increase in genomics knowledge and a positive perspective towards PGx. CONCLUSION: A course such as this bridges the current PGx basic science knowledge gap and can serve as a template for providing fundamental genomics knowledge until admissions prerequisites ensure that all incoming students are ready to embrace PGx.

Out-of-school genomics program for young women demystifies genomics and fosters interest in biomedical sciences.

BACKGROUND: Pharmacogenomics (PGx) plays a critical role in personalized medicine; however, most healthcare practitioners lack the training and confidence in PGx required to fully utilize its potential. Although continuing education and inclusion of PGx into the professional curricula will begin to address this deficiency, PGx education at the secondary and postsecondary levels would demystify PGx and pique interest at an academic stage that is key to funneling PGx curious students into the science and healthcare pipeline earlier. METHODS: This article describes the development and evaluation of a genomics outreach program targeted at young women in high school with the ultimate goal of recruiting students into an undergraduate PGx program and school of pharmacy. RESULTS: This program increased participants' genomics knowledge, influenced their careers interests, and imparted positive feelings towards a genomics-based and/or biomedical career. CONCLUSION: Genomics-based educational outreach programs geared towards secondary school students can increase interest in and confidence to pursue a PGx-centric degree/career. Original submitted 3 July 2013; Revision submitted 6 January 2014.

The current and future state of pharmacogenomics medical education in the USA.

Healthcare professionals (e.g., physicians, physician assistants, pharmacists, nurses and genetic counselors) believe pharmacogenomics (PGx) is essential to personalized medicine; however, they still lack confidence prescribing, dosing, interacting with other healthcare professionals and counseling patients with regard to PGx. This is due to the inadequate incorporation of PGx content into professional curricula. Compared with other health professions, Doctor of Pharmacy programs have integrated more PGx content. Unlike other healthcare professionals, pharmacists have extensive training in pharmacology, drug selection, drug dosage, drug-drug interactions and are uniquely accessible to patients. We suggest pharmacists are the best poised to facilitate incorporating PGx into therapeutic decision-making. Based on our experience as undergraduate and pharmacy PGx educators, we further reflect on our experience educating future healthcare professionals on PGx.

Dexamethasone-mediated repression of MUC5AC gene expression in human lung epithelial cells.

Glucocorticoids regulate gene expression via binding of the ligand-activated glucocorticoid receptor (GR) to glucocorticoid-responsive elements (GRE) in target gene promoters. The MUC5AC gene, which encodes the protein backbone of an abundant secreted airway mucin, has several putative GRE cis-elements in its 5' sequence. Mechanism(s) whereby glucocorticoids regulate mucin genes have not previously been described. In this study, the glucocorticoid dexamethasone (Dex) decreased MUC5AC mRNA abundance in A549 and NCI-H292 cell lines and primary differentiated normal bronchial epithelial cells by 50-80%, suggesting a common mechanism of MUC5AC gene repression in human lung epithelial cells. Kinetic analyses showed that MUC5AC mRNA was not significantly decreased until 6 h after Dex exposure, and that nuclear translocation of GR was biphasic, suggesting that Dex-mediated cis-repression of MUC5AC gene expression was a delayed response of GR translocation. Transfection analyses demonstrated that Dex transcriptionally repressed the MUC5AC promoter. Electrophoretic mobility shift assays with wild-type and mutant oligonucleotide probes showed that GR bound to two GRE cis-sites (nucleotides -930 to -912 and -369 to -351) in the MUC5AC promoter. Analyses of mutated MUC5AC promoter constructs demonstrated that NF-kappaB cis-sites were not involved in Dex-mediated repression of MUC5AC. Dex did not alter mRNA stability of MUC5AC transcripts. Taken together, the data indicate that Dex transcriptionally mediates repression of MUC5AC gene expression in human lung epithelial cells at quiescent states after binding of GR to one or more GRE cis-elements in the MUC5AC promoter.

Temporal analysis of goblet cells and mucin gene expression in murine models of allergic asthma.

In murine models of allergic asthma, mice repeatedly exposed to allergens or interleukin (IL)13 have numerous goblet cells in their airway epithelium, in contrast to healthy naïve mice. This study evaluated whether a single airway exposure of ovalbumin or IL13 would produce goblet cell metaplasia. Following ovalbumin challenge, airway goblet cells were present by 1 day, increased further by day 2 and day 3, and decreased by day 8. Following IL13 exposure, some goblet cells were detected at 6 hours and increased by 18 and 48 hours. Goblet transition cells, which are morphologically but not histologically similar to goblet cells, were observed at 6 and 18 hours following IL13 exposure and day 1 following ovalbumin challenge. Increased Muc5ac and Muc2 mRNA expression occurred following ovalbumin or IL13, but not saline, exposure. Mucin transcripts were localized to goblet cells in the surface airway epithelium. Muc5ac protein was expressed in some goblet transition and goblet cells. Overall, these data demonstrated that a single airway exposure to ovalbumin or IL13 is sufficient to generate goblet cell metaplasia and thus increase mucin gene expression in two strains of mice.