Function and Safety of Lentivirus-Mediated Gene Transfer for CSF2RA-Deficiency.

Autor:	Hetzel M; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Suzuki T; 3 Translational Pulmonary Science Center, Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.; 4 Department of Pediatrics, University of Cincinnati , Cincinnati, Ohio., Hashtchin AR; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Arumugam P; 3 Translational Pulmonary Science Center, Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.; 4 Department of Pediatrics, University of Cincinnati , Cincinnati, Ohio., Carey B; 3 Translational Pulmonary Science Center, Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.; 4 Department of Pediatrics, University of Cincinnati , Cincinnati, Ohio., Schwabbauer M; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Kuhn A; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Meyer J; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Schambach A; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany.; 5 Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School , Boston, Massachusetts., Van Der Loo J; 6 The Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania., Moritz T; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany., Trapnell BC; 3 Translational Pulmonary Science Center, Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.; 4 Department of Pediatrics, University of Cincinnati , Cincinnati, Ohio.; 7 Department of Medicine, University of Cincinnati , Cincinnati, Ohio., Lachmann N; 1 Institute of Experimental Hematology , Hannover Medical School, Hannover, Germany.; 2 REBIRTH Cluster of Excellence, Hannover, Germany.
Jazyk:	angličtina
Zdroj:	Human gene therapy methods [Hum Gene Ther Methods] 2017 Dec; Vol. 28 (6), pp. 318-329. Date of Electronic Publication: 2017 Aug 30.
DOI:	10.1089/hgtb.2017.092
Abstrakt:	Hereditary pulmonary alveolar proteinosis (hPAP) is a rare disorder of pulmonary surfactant accumulation and hypoxemic respiratory failure caused by mutations in CSF2RA (encoding the granulocyte/macrophage colony-stimulating factor [GM-CSF] receptor α-chain [CD116]), which results in reduced GM-CSF-dependent pulmonary surfactant clearance by alveolar macrophages. While no pharmacologic therapy currently exists for hPAP, it was recently demonstrated that endotracheal instillation of wild-type or gene-corrected mononuclear phagocytes (pulmonary macrophage transplantation [PMT]) results in a significant and durable therapeutic efficacy in a validated murine model of hPAP. To facilitate the translation of PMT therapy to human hPAP patients, a self-inactivating (SIN) lentiviral vector was generated expressing a codon-optimized human CSF2RA-cDNA driven from an EF1α short promoter (Lv.EFS.CSF2RA coop ), and a series of nonclinical efficacy and safety studies were performed in cultured macrophage cell lines and primary human cells. Studies in cytokine-dependent Ba/F3 cells demonstrated efficient transduction, vector-derived CD116 expression proportional to vector copy number, and GM-CSF-dependent cell survival and proliferation. Using a novel cell line constructed to express a normal GM-CSF receptor β subunit and a dysfunctional α subunit (due to a function-altering CSF2RA G196R mutation) that reflects the macrophage disease phenotype of hPAP patients, it was demonstrated that Lv.EFS.CSF2RA coop transduction restored GM-CSF receptor function. Further, Lv.EFS.CSF2RA coop transduction of healthy primary CD34 + cells did not adversely affect cell proliferation or affect the cell differentiation program. Results demonstrate Lv.EFS.CSF2RA coop reconstituted GM-CSF receptor α expression, restoring GM-CSF signaling in hPAP macrophages, and had no adverse effects in the intended target cells, thus supporting testing of PMT therapy of hPAP in humans.
Databáze:	MEDLINE
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