title: Gene editing of DNAH11 restores normal cilia motility in primary ciliary dyskinesia.
abstract: BACKGROUND
  Primary ciliary dyskinesia (PCD) is a rare autosomal recessive genetic disorder characterised by dysfunction of motile cilia. Ciliary dysmotility causes poor mucociliary clearance and leads to impairment of pulmonary function and severe respiratory infections. PCD has no specific therapy. With the aim to permanently restore gene function and normalise ciliary motility, we used gene editing to replace mutated with wild-type sequence in defective cells.
  
  
  METHODS
  The target gene was dynein heavy chain 11 (DNAH11), an essential component of ciliary structure. Airway ciliated cells were collected from two patients with PCD with DNAH11 nonsense mutations and altered ciliary beating and pattern. Repair of the genetic defect was performed ex vivo by site-specific recombination using transcription activator-like effector nucleases (TALENs).
  
  
  RESULTS
  In an epithelial cell line engineered to contain the DNAH11 target site, TALENs cleaved over 80% of the mutated DNAH11 sequence and replaced the mutated sequence with wild-type sequence in about 50% of cells. In airway ciliated cells of patients with PCD, site-specific recombination and normalisation of ciliary beating and pattern occurred in 33% and 29% of cells, respectively.
  
  
  CONCLUSIONS
  This study demonstrates that gene editing can rescue ciliary beating ex vivo, opening up new avenues for treating PCD.
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Lai
authors.affiliation: Department of Pediatrics, University Hospital of Pisa, Pisa, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Pifferi
authors.affiliation: Imperial College and Royal Brompton Hospital, London, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Bush
authors.affiliation: Department of Pediatrics, University Hospital of Pisa, Pisa, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Piras
authors.affiliation: Medical Genetics Laboratory, University Hospital of Pisa, Pisa, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Michelucci
authors.affiliation: Department of Pediatrics, University Hospital of Pisa, Pisa, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Di Cicco
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy.
authors.forename: Ambra
authors.initials: A
authors.lastname: del Grosso
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy ARPA Foundation, Pisa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Quaranta
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Cursi
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Tantillo
authors.affiliation: Fondazione Pisana per la Scienza, Pisa, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Franceschi
authors.affiliation: Fondazione Pisana per la Scienza, Pisa, Italy.
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Mazzanti
authors.affiliation: Medical Genetics Laboratory, University Hospital of Pisa, Pisa, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Simi
authors.affiliation: Department of Pediatrics, University Hospital of Pisa, Pisa, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Saggese
authors.affiliation: Department of Pediatrics, University of Verona, Verona, Italy.
authors.forename: Attilio
authors.initials: A
authors.lastname: Boner
authors.affiliation: Retrovirus Center and Virology Section, Department of Translational Research, University of Pisa, Pisa, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Pistello
mesh_terms: D000293:Adolescent; D056444:Axonemal Dyneins / Q000235:genetics*; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics; D002923:Cilia / Q000378:metabolism / Q000473:pathology; D004847:Epithelial Cells / Q000473:pathology; D000072669:Gene Editing; D015316:Genetic Therapy; D005838:Genotype; D006801:Humans; D007619:Kartagener Syndrome / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D016086:Lentivirus / Q000235:genetics; D008297:Male; D010641:Phenotype; D014427:Twins
chemical_list: D056444:Axonemal Dyneins; C465433:DNAH11 protein, human
keywords: Airway epithelial cells; Gene editing; PCD; Primary ciliary diskynesia; TALEN