@article{989654b76fef41ac99a9983384452dbf,
title = "Lack of GAS2L2 Causes PCD by Impairing Cilia Orientation and Mucociliary Clearance",
abstract = " Primary ciliary dyskinesia (PCD) is a genetic disorder in which impaired ciliary function leads to chronic airway disease. Exome sequencing of a PCD subject identified an apparent homozygous frameshift variant, c.887_890delTAAG (p.Val296Glyfs ∗ 13), in exon 5; this frameshift introduces a stop codon in amino acid 308 of the growth arrest-specific protein 2-like 2 (GAS2L2). Further genetic screening of unrelated PCD subjects identified a second proband with a compound heterozygous variant carrying the identical frameshift variant and a large deletion (c.867_ ∗ 343+1207del; p.?) starting in exon 5. Both individuals had clinical features of PCD but normal ciliary axoneme structure. In this research, using human nasal cells, mouse models, and X.laevis embryos, we show that GAS2L2 is abundant at the apical surface of ciliated cells, where it localizes with basal bodies, basal feet, rootlets, and actin filaments. Cultured GAS2L2-deficient nasal epithelial cells from one of the affected individuals showed defects in ciliary orientation and had an asynchronous and hyperkinetic (GAS2L2-deficient = 19.8 Hz versus control = 15.8 Hz) ciliary-beat pattern. These results were recapitulated in Gas2l2 −/− mouse tracheal epithelial cell (mTEC) cultures and in X. laevis embryos treated with Gas2l2 morpholinos. In mice, the absence of Gas2l2 caused neonatal death, and the conditional deletion of Gas2l2 impaired mucociliary clearance (MCC) and led to mucus accumulation. These results show that a pathogenic variant in GAS2L2 causes a genetic defect in ciliary orientation and impairs MCC and results in PCD.",
keywords = "GAS2L2, MCC, PCD, ciliary orientation, mucociliary clearance, primary ciliary dyskinesia",
author = "Bustamante-Marin, {Ximena M.} and Yin, {Wei Ning} and Sears, {Patrick R.} and Werner, {Michael E.} and Brotslaw, {Eva J.} and Mitchell, {Brian J.} and Jania, {Corey M.} and Zeman, {Kirby L.} and Rogers, {Troy D.} and Herring, {Laura E.} and Luc Refab{\'e}rt and Lucie Thomas and Serge Amselem and Estelle Escudier and Marie Legendre and Grubb, {Barbara R.} and Knowles, {Michael R.} and Zariwala, {Maimoona A.} and Ostrowski, {Lawrence E.}",
note = "Funding Information: The authors would like to thank the PCD subjects and family members for their participation and thank the US PCD Foundation and the investigators and coordinators of the Genetic Disorders of Mucociliary Clearance Consortium, part of the Rare Disease Clinical Research Network. We thank Jay Shendure, Deborah Nickerson, and Michael Bamshad from University of Washington School of Medicine (National Institutes of Health [NHI]/ National Human Genome Research Institute [NHGRI] U54HG0006493), Seattle GO Sequencing Project (HL-102926), RS & G at Northwest Genomics Center at the University of Washington (NIH-National Heart, Lung, and Blood Institute [NHLBI] HHSN268201100037); Shrikant Mane, Francesc Lopez-Giraldez, and Weilai Dong from the Yale Center (UM1 HG006504) for whole-exome sequencing and bioinformatics support; Kimberly Burns for histology and electron-microscopy support; Whitney Wolf and Lu Huang for technical assistance; Hong Dang for bioinformatics assistance; Robert Tarran for providing the Leica Sp8 confocal and GSD microscopes; Michael Chua and Tony Perdue for assistance with confocal imaging; Frank Conlon and Panna Tandon for providing Xenopus laevis oocytes; Uma Nagarajan and Amy Kiatthanapaiboon for providing samples of human fallopian-tube total RNA; Julie Kimbell for assistance with Mimics Research 18.0; and Scott H. Randell and the UNC Cell Culture Core for providing human airway cells. The UNC Cell Core Facility is supported by BOUCHE15R0 and P30DK065988 grants. The UNC Proteomics Core Facility is supported in part by P30 CA016086. The Small Animal Imaging Facility at the UNC Biomedical Imaging Research Center is supported in part by a National Cancer Institute cancer core grant, P30-CA016086-40. Funding support for this research was provided to B.J.M. by R01GM089970 from the National Institute of General Medical Sciences at the NIH; to M.R.K and M.A.Z by grant 5U54HL096458 from the Office of Rare Diseases Research and NHLBI at the NIH; to M.R.K., L.E.O., and M.A.Z. by NIH-NHLBI grant R01HL071798; to L.E.O., M.A.Z., and M.R.K by NIH-NHLBI grant R01HL117836; and to the University of North Carolina at Chapel Hill by grant UL1 TR000083 from the National Center for Advancing Translational Sciences at the NIH. Work by S.A., E.E., M.L., and L.T. is supported by the Fondation pour la Recherche M{\'e}dicale grant DEQ20120323689, by Institut National de la Sant{\'e} et de la Recherche M{\'e}dicale (INSERM) grant ANR-10-COHO-003, and the Legs Poix grant from the Chancellerie des Universit{\'e}s of Sorbonne Universit{\'e}. Funding Information: The authors would like to thank the PCD subjects and family members for their participation and thank the US PCD Foundation and the investigators and coordinators of the Genetic Disorders of Mucociliary Clearance Consortium, part of the Rare Disease Clinical Research Network. We thank Jay Shendure, Deborah Nickerson, and Michael Bamshad from University of Washington School of Medicine ( National Institutes of Health [NHI]/ National Human Genome Research Institute [NHGRI] U54HG0006493 ), Seattle GO Sequencing Project ( HL-102926 ), RS & G at Northwest Genomics Center at the University of Washington ( NIH-National Heart, Lung, and Blood Institute [NHLBI] HHSN268201100037 ); Shrikant Mane, Francesc Lopez-Giraldez, and Weilai Dong from the Yale Center ( UM1 HG006504 ) for whole-exome sequencing and bioinformatics support; Kimberly Burns for histology and electron-microscopy support; Whitney Wolf and Lu Huang for technical assistance; Hong Dang for bioinformatics assistance; Robert Tarran for providing the Leica Sp8 confocal and GSD microscopes; Michael Chua and Tony Perdue for assistance with confocal imaging; Frank Conlon and Panna Tandon for providing Xenopus laevis oocytes; Uma Nagarajan and Amy Kiatthanapaiboon for providing samples of human fallopian-tube total RNA; Julie Kimbell for assistance with Mimics Research 18.0; and Scott H. Randell and the UNC Cell Culture Core for providing human airway cells. The UNC Cell Core Facility is supported by BOUCHE15R0 and P30DK065988 grants. The UNC Proteomics Core Facility is supported in part by P30 CA016086 . The Small Animal Imaging Facility at the UNC Biomedical Imaging Research Center is supported in part by a National Cancer Institute cancer core grant, P30-CA016086-40 . Funding support for this research was provided to B.J.M. by R01GM089970 from the National Institute of General Medical Sciences at the NIH ; to M.R.K and M.A.Z by grant 5U54HL096458 from the Office of Rare Diseases Research and NHLBI at the NIH ; to M.R.K., L.E.O., and M.A.Z. by NIH-NHLBI grant R01HL071798 ; to L.E.O., M.A.Z., and M.R.K by NIH-NHLBI grant R01HL117836 ; and to the University of North Carolina at Chapel Hill by grant UL1 TR000083 from the National Center for Advancing Translational Sciences at the NIH . Work by S.A., E.E., M.L., and L.T. is supported by the Fondation pour la Recherche M{\'e}dicale grant DEQ20120323689 , by Institut National de la Sant{\'e} et de la Recherche M{\'e}dicale (INSERM) grant ANR-10-COHO-003 , and the Legs Poix grant from the Chancellerie des Universit{\'e}s of Sorbonne Universit{\'e} . Publisher Copyright: {\textcopyright} 2018 American Society of Human Genetics",
year = "2019",
month = feb,
day = "7",
doi = "10.1016/j.ajhg.2018.12.009",
language = "English (US)",
volume = "104",
pages = "229--245",
journal = "American Journal of Human Genetics",
issn = "0002-9297",
publisher = "Cell Press",
number = "2",
}
