Lack of airway submucosal glands impairs respiratory host defenses

Ostedgaard, Lynda S.; Price, Margaret P.; Whitworth, Kristin M.; Alaiwa, Mahmoud H. Abou; Fischer, Anthony J.; Warrier, Akshaya; Samuel, Melissa; Spate, Lee D.; Allen, Patrick D.; Hilkin, Brieanna M.; Ibarra, Guillermo S. Romano; Bezara, Miguel E Ortiz; Goodell, Brian J; Mather, Steven E; Powers, Linda S.; Stroik, Mallory R.; Gansemer, Nicholas D.; Hippee, Camilla E; Zarei, Keyan; Goeken, J. Adam; Businga, Thomas R.; Hoffman, Eric A.; Meyerholz, David K.; Prather, Randall S.; Stoltz, David A.; Welsh, Michael J.

doi:10.7554/elife.59653

Cited by 28 publications

(30 citation statements)

References 103 publications

(213 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The recent generation of a genetically modified pig model lacking SMG (EDA-KO pig; ref. 66 ) highlights how critical these studies are for our understanding of CF pathophysiology. In this context, the CF rabbit offers a suitable model to study the effect of CFTR functional depletion in airway compartments that are normally devoid of SMG (trachea and lungs) or containing SMG (nose and nasopharynx).…”

Section: Discussionmentioning

confidence: 99%

Phenotypes of CF rabbits generated by CRISPR/Cas9-mediated disruption of the CFTR gene

Xu¹,

Livraghi‐Butrico²,

Hou³

et al. 2021

JCI Insight

View full text Add to dashboard Cite

Existing animal models of cystic fibrosis (CF) have provided key insights into CF pathogenesis but have been limited by short life spans, absence of key phenotypes, and/or high maintenance costs. Here, we report the CRISPR/Cas9-mediated generation of CF rabbits, a model with a relatively long lifespan and affordable maintenance and care costs. CF rabbits supplemented solely with oral osmotic laxative had a median survival of ~ 40 days and died of gastrointestinal disease, but therapeutic regimens directed toward restoring gastrointestinal transit extended median survival to ~ 80 days. Surrogate markers of exocrine pancreas disorders were found in CF rabbits with declining health. CFTR expression patterns in WT rabbit airways mimicked humans, with widespread distribution in nasal respiratory and olfactory epithelia, as well as proximal and distal lower airways. CF rabbits exhibited human CF-like abnormalities in the bioelectric properties of the nasal and tracheal epithelia. No spontaneous respiratory disease was detected in young CF rabbits. However, abnormal phenotypes were observed in surviving 1 year-old CF rabbits as compared to WT littermates, which were especially evident in the nasal respiratory and olfactory epithelium. The CF rabbit model may serve as a useful tool for understanding gut and lung CF pathogenesis and for the practical development of CF therapeutics.

show abstract

Section: Discussionmentioning

confidence: 99%

Phenotypes of CF rabbits generated by CRISPR/Cas9-mediated disruption of the CFTR gene

Xu¹,

Livraghi‐Butrico²,

Hou³

et al. 2021

JCI Insight

View full text Add to dashboard Cite

show abstract

“…Other relevant oxidative stress-sensitive aspects intrinsic of CF airway cell physiology that appeared significantly highlighted in our pathway analysis refer to cell remodeling, cell-cell communication, and motility. It has been recently shown that submucosal glands are required for normal antimicrobial activity and mucociliary transport, two key host defenses protecting the lung that are disabled in CF patients [ 69 ]. Thus, it is not casual that related pathways such as intraflagellar transport and cilium assembly are significantly overrepresented in our study, because they may impact airway epithelial cell integrity upon repeated oxidative stress challenges experienced by CF patients.…”

Section: Discussionmentioning

confidence: 99%

Genome-Wide RNAi Screening Identifies Novel Pathways/Genes Involved in Oxidative Stress and Repurposable Drugs to Preserve Cystic Fibrosis Airway Epithelial Cell Integrity

et al. 2021

View full text Add to dashboard Cite

Recurrent infection-inflammation cycles in cystic fibrosis (CF) patients generate a highly oxidative environment, leading to progressive destruction of the airway epithelia. The identification of novel modifier genes involved in oxidative stress susceptibility in the CF airways might contribute to devise new therapeutic approaches. We performed an unbiased genome-wide RNAi screen using a randomized siRNA library to identify oxidative stress modulators in CF airway epithelial cells. We monitored changes in cell viability after a lethal dose of hydrogen peroxide. Local similarity and protein-protein interaction network analyses uncovered siRNA target genes/pathways involved in oxidative stress. Further mining against public drug databases allowed identifying and validating commercially available drugs conferring oxidative stress resistance. Accordingly, a catalog of 167 siRNAs able to confer oxidative stress resistance in CF submucosal gland cells targeted 444 host genes and multiple circuitries involved in oxidative stress. The most significant processes were related to alternative splicing and cell communication, motility, and remodeling (impacting cilia structure/function, and cell guidance complexes). Other relevant pathways included DNA repair and PI3K/AKT/mTOR signaling. The mTOR inhibitor everolimus, the α1-adrenergic receptor antagonist doxazosin, and the Syk inhibitor fostamatinib significantly increased the viability of CF submucosal gland cells under strong oxidative stress pressure. Thus, novel therapeutic strategies to preserve airway cell integrity from the harsh oxidative milieu of CF airways could stem from a deep understanding of the complex consequences of oxidative stress at the molecular level, followed by a rational repurposing of existing “protective” drugs. This approach could also prove useful to other respiratory pathologies.

show abstract

“…In the mouse, MUC5AC contributes to airway hyperreactivity (Evans et al, 2015) and mucus plugging, whereas MUC5B is required for airway homeostasis, mucociliary clearance, and anti‐bacterial defenses (Roy et al, 2014). In the pig, elimination of airway submucosal glands, which predominantly secrete MUC5B, also impairs mucociliary clearance and airway host defenses (Ostedgaard et al, 2020). We found that overexpression of SP selectively increased MUC5AC.…”

Section: Discussionmentioning

confidence: 99%

Overexpression of Substance P in pig airways increases MUC5AC through an NF‐kβ pathway

et al. 2021

View full text Add to dashboard Cite

Substance P (SP) is a tachykinin that regulates airway mucous secretion in both health and disease. Our study aimed to determine whether overexpression of SP without pre‐existing inflammation was sufficient to induce changes in mucin secretion and transport in small airways. Utilizing porcine precision‐cut lung slices, we measured the impact of AAV‐mediated overexpression of SP on airway physiology ex vivo. Immunofluorescence signal intensity for MUC5AC was significantly increased in SP‐overexpressed precision‐cut lung slices compared to GFP controls. No difference in MUC5B signal intensity between treatments was detected. SP‐overexpressed precision‐cut lung slices also exhibited decreased IL10 mRNA, an important inhibitor of mucous cell metaplasia. Overt deficits in mucociliary transport were not noted, though a trend for decreased mean transport speed was detected in methacholine‐challenged airways overexpressing SP compared to GFP controls. Pharmacologic inhibition of the NF‐kβ pathway abrogated the effects of overexpression of SP on both MUC5AC and IL10. Collectively, these data suggest that overexpression of SP in the absence of existing inflammation increases MUC5AC via activation of the NF‐kβ pathway. Thus, these data further highlight SP as a key driver of abnormal mucous secretion and underscore NF‐kβ signaling as a pathway of potential therapeutic intervention.

show abstract

Lack of airway submucosal glands impairs respiratory host defenses

Cited by 28 publications

References 103 publications

Phenotypes of CF rabbits generated by CRISPR/Cas9-mediated disruption of the CFTR gene

Phenotypes of CF rabbits generated by CRISPR/Cas9-mediated disruption of the CFTR gene

Genome-Wide RNAi Screening Identifies Novel Pathways/Genes Involved in Oxidative Stress and Repurposable Drugs to Preserve Cystic Fibrosis Airway Epithelial Cell Integrity

Overexpression of Substance P in pig airways increases MUC5AC through an NF‐kβ pathway

Contact Info

Product

Resources

About