TY - JOUR
T1 - Transcriptional analysis of primary ciliary dyskinesia airway cells reveals a dedicated cilia glutathione pathway
AU - Koenitzer, Jeffrey R.
AU - Gupta, Deepesh Kumar
AU - Twan, Wang Kyaw
AU - Xu, Huihui
AU - Hadas, Nicholas
AU - Hawkins, Finn J.
AU - Beermann, Mary Lou
AU - Penny, Gervette M.
AU - Wamsley, Nathan T.
AU - Berical, Andrew
AU - Major, Michael B.
AU - Dutcher, Susan K.
AU - Brody, Steven L.
AU - Horani, Amjad
N1 - Publisher Copyright:
© 2024, Koenitzer et al.
PY - 2024/9/10
Y1 - 2024/9/10
N2 - Primary ciliary dyskinesia (PCD) is a genetic condition that results in dysmotile cilia. The repercussions of cilia dysmotility and gene variants on the multiciliated cell remain poorly understood. We used single-cell RNA-Seq, proteomics, and advanced microscopy to compare primary culture epithelial cells from patients with PCD, their heterozygous mothers, and healthy individuals, and we induced pluripotent stem cells (iPScs) generated from a patient with PCD. Transcriptomic analysis revealed unique signatures in PCD airway cells compared with their mothers’ cells and the cells of healthy individuals. Gene expression in heterozygous mothers’ cells diverged from both control and PCD cells, marked by increased inflammatory and cellular stress signatures. Primary and iPS-derived PCD multiciliated cells had increased expression of glutathione-S-transferases GSTA2 and GSTA1, as well as NRF2 target genes, accompanied by elevated levels of reactive oxygen species (ROS). Immunogold labeling in human cilia and proteomic analysis of the ciliated organism Chlamydomonas reinhardtii demonstrated that GSTA2 localizes to motile cilia. Loss of human GSTA2 and C. reinhardtii GSTA resulted in slowed cilia motility, pointing to local cilia regulatory roles. Our findings identify cellular responses unique to PCD variants and independent of environmental stress and uncover a dedicated ciliary GSTA2 pathway essential for normal motility that may be a therapeutic target.
AB - Primary ciliary dyskinesia (PCD) is a genetic condition that results in dysmotile cilia. The repercussions of cilia dysmotility and gene variants on the multiciliated cell remain poorly understood. We used single-cell RNA-Seq, proteomics, and advanced microscopy to compare primary culture epithelial cells from patients with PCD, their heterozygous mothers, and healthy individuals, and we induced pluripotent stem cells (iPScs) generated from a patient with PCD. Transcriptomic analysis revealed unique signatures in PCD airway cells compared with their mothers’ cells and the cells of healthy individuals. Gene expression in heterozygous mothers’ cells diverged from both control and PCD cells, marked by increased inflammatory and cellular stress signatures. Primary and iPS-derived PCD multiciliated cells had increased expression of glutathione-S-transferases GSTA2 and GSTA1, as well as NRF2 target genes, accompanied by elevated levels of reactive oxygen species (ROS). Immunogold labeling in human cilia and proteomic analysis of the ciliated organism Chlamydomonas reinhardtii demonstrated that GSTA2 localizes to motile cilia. Loss of human GSTA2 and C. reinhardtii GSTA resulted in slowed cilia motility, pointing to local cilia regulatory roles. Our findings identify cellular responses unique to PCD variants and independent of environmental stress and uncover a dedicated ciliary GSTA2 pathway essential for normal motility that may be a therapeutic target.
UR - http://www.scopus.com/inward/record.url?scp=85203857297&partnerID=8YFLogxK
U2 - 10.1172/jci.insight.180198
DO - 10.1172/jci.insight.180198
M3 - Article
C2 - 39042459
AN - SCOPUS:85203857297
SN - 2379-3708
VL - 9
JO - JCI Insight
JF - JCI Insight
IS - 17
M1 - e180198
ER -