title: Gene correction of human induced pluripotent stem cells repairs the cellular phenotype in pulmonary alveolar proteinosis.
abstract: BACKGROUND
  Hereditary pulmonary alveolar proteinosis (hPAP) caused by granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor α-chain (CSF2RA) deficiency is a rare, life-threatening lung disease characterized by accumulation of proteins and phospholipids in the alveolar spaces. The disease is caused by a functional insufficiency of alveolar macrophages, which require GM-CSF signaling for terminal differentiation and effective degradation of alveolar proteins and phospholipids. Therapeutic options are extremely limited, and the pathophysiology underlying the defective protein degradation in hPAP alveolar macrophages remains poorly understood.
  
  
  OBJECTIVE
  To further elucidate the cellular mechanisms underlying hPAP and evaluate novel therapeutic strategies, we here investigated the potential of hPAP patient-derived induced pluripotent stem cell (PAP-iPSCs) derived monocytes and macrophages.
  
  
  METHODS
  Patient-specific PAP-iPSCs were generated from CD34(+) bone marrow cells of a CSF2RA-deficient patient with PAP. We assessed pluripotency, chromosomal integrity, and genetic correction of established iPSC lines. On hematopoietic differentiation, genetically corrected or noncorrected monocytes and macrophages were investigated in GM-CSF-dependent assays.
  
  
  RESULTS
  Although monocytes and macrophages differentiated from noncorrected PAP-iPSCs exhibited distinct defects in GM-CSF-dependent functions, such as perturbed CD11b activation, phagocytic activity, and STAT5 phosphorylation after GM-CSF exposure and lack of GM-CSF uptake, these defects were fully repaired on lentiviral gene transfer of a codon-optimized CSF2RA-cDNA.
  
  
  CONCLUSIONS
  These data establish PAP-iPSC-derived monocytes and macrophages as a valid in vitro disease model of CSF2RA-deficient PAP, and introduce gene-corrected iPSC-derived monocytes and macrophages as a potential autologous cell source for innovative therapeutic strategies. Transplantation of such cells to patients with hPAP could serve as a paradigmatic proof for the potential of iPSC-derived cells in clinical gene therapy.
authors.affiliation: 1 Reprogramming and Gene Therapy Group, REBIRTH Cluster of Excellence.
authors.forename: Nico
authors.initials: N
authors.lastname: Lachmann
authors.forename: Christine
authors.initials: C
authors.lastname: Happle
authors.forename: Mania
authors.initials: M
authors.lastname: Ackermann
authors.forename: Doreen
authors.initials: D
authors.lastname: Lüttge
authors.forename: Martin
authors.initials: M
authors.lastname: Wetzke
authors.forename: Sylvia
authors.initials: S
authors.lastname: Merkert
authors.forename: Miriam
authors.initials: M
authors.lastname: Hetzel
authors.forename: George
authors.initials: G
authors.lastname: Kensah
authors.forename: Monica
authors.initials: M
authors.lastname: Jara-Avaca
authors.forename: Adele
authors.initials: A
authors.lastname: Mucci
authors.forename: Jelena
authors.initials: J
authors.lastname: Skuljec
authors.forename: Anna-Maria
authors.initials: AM
authors.lastname: Dittrich
authors.forename: Nils
authors.initials: N
authors.lastname: Pfaff
authors.forename: Sebastian
authors.initials: S
authors.lastname: Brennig
authors.forename: Axel
authors.initials: A
authors.lastname: Schambach
authors.forename: Doris
authors.initials: D
authors.lastname: Steinemann
authors.forename: Gudrun
authors.initials: G
authors.lastname: Göhring
authors.forename: Tobias
authors.initials: T
authors.lastname: Cantz
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
authors.forename: Nicolaus
authors.initials: N
authors.lastname: Schwerk
authors.forename: Gesine
authors.initials: G
authors.lastname: Hansen
authors.forename: Thomas
authors.initials: T
authors.lastname: Moritz
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002675:Child, Preschool; D005260:Female; D040181:Genetic Diseases, X-Linked / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D015316:Genetic Therapy; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D016676:Macrophages, Alveolar / Q000378:metabolism; D008954:Models, Biological; D009000:Monocytes / Q000378:metabolism; D011649:Pulmonary Alveolar Proteinosis / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000172:deficiency* / Q000235:genetics / Q000627:therapeutic use; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
chemical_list: C521678:CSF2RA protein, human; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor