Highlights
- •In the lung, Orai1 expression is highest in immune cells.
- •Orai1/STIM1 colocalization is increased in asthma and CF lung epithelial, interstitial, and luminal immune cells.
- •Orai1 aggregation and activation is increased in asthmatic and CF lung immune cells.
- •Orai1 activation is elevated in CF patient blood neutrophils regardless of CFTR modulator usage.
- •Orai1 is a relevant target for reducing pulmonary inflammation.
Abstract
Question
In diseases such as asthma and cystic fibrosis (CF), the immune response is dysregulated
and the lung is chronically inflamed. Orai1 activation is required for the initiation
and persistence of inflammation. However, Orai1 expression in the lung is poorly understood.
We therefore tested the hypothesis that Orai1 expression was upregulated in asthmatic
and CF lungs.
Materials and methods
We used LungMAP to analyze single-cell RNAseq data of Orai1 and stromal interaction
molecule 1 (STIM1) expression in normal human lungs. We then performed RNAscope analysis
and immunostaining on lung sections from normal, asthma, and CF donors. We imaged
sections by confocal and super resolution microscopy, and analyzed Orai1 and STIM1
expression in different pulmonary cell types.
Results
Orai1 was broadly-expressed, but expression was greatest in immune cells. At mRNA
and protein levels, there were no consistent trends in expression levels between the
three phenotypes. Orai1 must interact with STIM1 in order to activate and conduct
Ca2+. We therefore used STIM1/Orai1 co-localization as a marker of Orai1 activity. Using
this approach, we found significantly increased co-localization between these proteins
in epithelia, interstitial and luminal immune cells, but not alveoli, from asthma
and CF lungs. Orai1 also aggregates as part of its activation process. Using super
resolution microscopy, we also found significantly increased Orai1 aggregation in
immune cells from asthmatic and CF lungs.
Conclusion
We found evidence that Orai1 was more active in asthma and CF than normal lungs. These
data suggest that Orai1 is a relevant target for reducing pulmonary inflammation.
Keywords
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Article info
Publication history
Published online: August 10, 2022
Accepted:
July 4,
2022
Received in revised form:
July 1,
2022
Received:
March 9,
2022
Identification
Copyright
© 2022 Published by Elsevier B.V. on behalf of European Cystic Fibrosis Society.
