| Autor: |
Koenitzer JR; Department of Medicine and., Gupta DK; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA., Twan WK; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA., Xu H; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA., Hadas N; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA., Hawkins FJ; Center for Regenerative Medicine and.; The Pulmonary Center, Department of Medicine, Boston University and Boston Medical Center, Boston, Massachusetts, USA., Beermann ML; Center for Regenerative Medicine and., Penny GM; Department of Genetics and., Wamsley NT; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, USA., Berical A; Center for Regenerative Medicine and.; The Pulmonary Center, Department of Medicine, Boston University and Boston Medical Center, Boston, Massachusetts, USA., Major MB; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, USA., Dutcher SK; Department of Genetics and.; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, USA., Brody SL; Department of Medicine and., Horani A; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA.; Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri, USA. |
| Abstrakt: |
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. |
