XHigh-content siRNA screen reveals global ENaC regulators and potential cystic fibrosis therapy targets

Joana Almaça, Diana Faria, Marisa Sousa, Inna Uliyakina, Christian Conrad, Lalida Sirianant, Luka A. Clarke, José Paulo Martins, Miguel Santos, Jean Karim Heriché, Wolfgang Huber, Rainer Schreiber, Rainer Pepperkok, Karl Kunzelmann, Margarida D. Amaral

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

Dysfunction of ENaC, the epithelial sodium channel that regulates salt and water reabsorption in epithelia, causes several human diseases, including cystic fibrosis (CF). To develop a global understanding of molecular regulators of ENaC traffic/function and to identify of candidate CF drug targets, we performed a large-scale screen combining high-content live-cell microscopy and siRNAs in human airway epithelial cells. Screening over 6,000 genes identified over 1,500 candidates, evenly divided between channel inhibitors and activators. Genes in the phosphatidylinositol pathway were enriched on the primary candidate list, and these, along with other ENaC activators, were examined further with secondary siRNA validation. Subsequent detailed investigation revealed ciliary neurotrophic factor receptor (CNTFR) as an ENaC modulator and showed that inhibition of (diacylglycerol kinase, iota) DGKι, a protein involved in PiP2 metabolism, downgrades ENaC activity, leading to normalization of both Na+ and fluid absorption in CF airways to non-CF levels in primary human lung cells from CF patients.

Original languageEnglish (US)
Number of pages1
JournalCell
Volume154
Issue number6
DOIs
StatePublished - Sep 12 2013
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Almaça, J., Faria, D., Sousa, M., Uliyakina, I., Conrad, C., Sirianant, L., Clarke, L. A., Martins, J. P., Santos, M., Heriché, J. K., Huber, W., Schreiber, R., Pepperkok, R., Kunzelmann, K., & Amaral, M. D. (2013). XHigh-content siRNA screen reveals global ENaC regulators and potential cystic fibrosis therapy targets. Cell, 154(6). https://doi.org/10.1016/j.cell.2013.08.045