• Nico Lachmann, Christine Happle, Mania Ackermann, Doreen Lüttge, Martin Wetzke, Sylvia Merkert, Miriam Hetzel, George Kensah, Monica Jara-Avaca, Adele Mucci, Jelena Skuljec, Anna-Maria Dittrich, Nils Pfaff, Sebastian Brennig, Axel Schambach, Doris Steinemann, Gudrun Göhring, Tobias Cantz, Ulrich Martin, Nicolaus Schwerk, Gesine Hansen, and Thomas Moritz.
• 1 Reprogramming and Gene Therapy Group, REBIRTH Cluster of Excellence.
• Am. J. Respir. Crit. Care Med.. 2014 Jan 15;189(2):167-82.

RationaleHereditary 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.ObjectivesTo 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.MethodsPatient-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 ResultsAlthough 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.ConclusionsThese 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.

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