RATIONALE: 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. OBJECTIVES: 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. MEASUREMENTS AND MAIN 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.
RATIONALE: 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. OBJECTIVES: To further elucidate the cellular mechanisms underlying hPAP and evaluate novel therapeutic strategies, we here investigated the potential of hPAPpatient-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. MEASUREMENTS AND MAIN 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: Tatsuya Morishima; Ann-Christin Krahl; Masoud Nasri; Yun Xu; Narges Aghaallaei; Betül Findik; Maksim Klimiankou; Malte Ritter; Marcus D Hartmann; Christian Johannes Gloeckner; Sylwia Stefanczyk; Christian Lindner; Benedikt Oswald; Regine Bernhard; Karin Hähnel; Ursula Hermanutz-Klein; Martin Ebinger; Rupert Handgretinger; Nicolas Casadei; Karl Welte; Maya Andre; Patrick Müller; Baubak Bajoghli; Julia Skokowa Journal: Blood Date: 2019-07-31 Impact factor: 22.113
Authors: D Hoffmann; J W Schott; F K Geis; L Lange; F-J Müller; D Lenz; D Zychlinski; D Steinemann; M Morgan; T Moritz; A Schambach Journal: Gene Ther Date: 2017-04-20 Impact factor: 5.250
Authors: Arnold S Kristof; Basil J Petrof; Qutayba Hamid; Martin Kolb; Jennifer S Landry; Alex MacKenzie; Francis X McCormack; Inga J Murawski; Joel Moss; Frank Rauch; Ivan O Rosas; Adam J Shapiro; Benjamin M Smith; David Y Thomas; Bruce C Trapnell; Lisa R Young; Maimoona A Zariwala Journal: Ann Am Thorac Soc Date: 2017-08
Authors: Miriam Hetzel; Takuji Suzuki; Anna Rafiei Hashtchin; Paritha Arumugam; Brenna Carey; Marc Schwabbauer; Alexandra Kuhn; Johann Meyer; Axel Schambach; Johannes Van Der Loo; Thomas Moritz; Bruce C Trapnell; Nico Lachmann Journal: Hum Gene Ther Methods Date: 2017-08-30 Impact factor: 2.396