Gene Correction of Human Induced Pluripotent Stem Cells Repairs the Cellular Phenotype in Pulmonary Alveolar Proteinosis

Abstract

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.

Ex vivo Generation of Genetically Modified Macrophages from Human Induced Pluripotent Stem Cells

“…In order to avoid silencing of the GFP transgene, we 1C). Similar results could be obtained for a second iPSC line (CD34iPSC16) [13]. Here lentiviral transduction with a MOI of 50 resulted in >90% stable GFP expression and maintained iPSC morphology ( fig.…”

“…Although our initial studies used HSPCs derived from murine bone marrow or human cord blood, also iPSC-derived macrophages represent a highly interesting source for cell therapy strategies in diseases related to macrophage dysfunction [13,14]. Here, the embryonic origin of the cells may even serve as an advantage [15], and the transplantation of iPSC-derived macrophages potentially may be superior to the transplantation of adult CD34+ HSC-derived macrophages.…”

“…Whereas our initial proof-of-concept studies focused on the expression of fluorescent reporter proteins, the A2UCOE has also been applied efficiently in the context of iPSC-based gene therapy for hereditary pulmonary alveolar proteinosis. Utilizing patient-specific iPSCs, an A2UCOE-equipped lentiviral vector was demonstrated to restore robust and stable expression of the GM-CSF receptor α-chain in hiPSCs and thereof derived macrophages [13]. In order to further optimize the A2U-COE, we recently generated a minimal 0.7 kb CBX3-UCOE [24].…”

Preclinical safety and efficacy models for pulmonary drug delivery of antimicrobials with focus on in vitro models

“…Therefore, in vitro models can be quite useful. Bueno emphasizes that such models save time and money during drug screening, especially in the early stages of research [136]. He suggested a stepby-step procedure to identify possible candidates (Fig.…”

Update on Diffuse Lung Disease in Children

“…In csf2rb -/-mice that have an analogous disease to human PAP, a macrophage transplantation consisting of either wild-type murine macrophages, gene-corrected macrophages from mutant mice, or normal human macrophages resulted in significantly improved surfactant clearance and homeostasis. [65][66][67] Furthermore, the effects lasted for up to 9 months, longer than the average life span of a pulmonary macrophage, indicating that a normal population of pulmonary macrophages can be established in the lungs of the affected mice. If these findings can be recapitulated in humans, they would significantly improve therapy for this form of PAP and could potentially be curative.…”