TY - JOUR
T1 - Safe and efficient gene therapy for pyruvate kinase deficiency
AU - Garcia-Gomez, Maria
AU - Calabria, Andrea
AU - Garcia-Bravo, Maria
AU - Benedicenti, Fabrizio
AU - Kosinski, Penelope
AU - López-Manzaneda, Sergio
AU - Hill, Collin
AU - Del Mar Mau-Pereira, María
AU - Martín, Miguel A.
AU - Orman, Israel
AU - Vives-Corrons, Joan Lluis
AU - Kung, Charles
AU - Schambach, Axel
AU - Jin, Shengfang
AU - Bueren, Juan A.
AU - Montini, Eugenio
AU - Navarro, Susana
AU - Segovia, Jose C.
PY - 2016/8/1
Y1 - 2016/8/1
N2 - Pyruvate kinase deficiency (PKD) is a monogenic metabolic disease caused by mutations in the PKLR gene that leads to hemolytic anemia of variable symptomatology and that can be fatal during the neonatal period. PKD recessive inheritance trait and its curative treatment by allogeneic bone marrow transplantation provide an ideal scenario for developing gene therapy approaches. Here, we provide a preclinical gene therapy for PKD based on a lentiviral vector harboring the hPGK eukaryotic promoter that drives the expression of the PKLR cDNA. This therapeutic vector was used to transduce mouse PKD hematopoietic stem cells (HSCs) that were subsequently transplanted into myeloablated PKD mice. Ectopic RPK expression normalized the erythroid compartment correcting the hematological phenotype and reverting organ pathology. Metabolomic studies demonstrated functional correction of the glycolytic pathway in RBCs derived from genetically corrected PKD HSCs, with no metabolic disturbances in leukocytes. The analysis of the lentiviral insertion sites in the genome of transplanted hematopoietic cells demonstrated no evidence of genotoxicity in any of the transplanted animals. Overall, our results underscore the therapeutic potential of the hPGK-coRPK lentiviral vector and provide high expectations toward the gene therapy of PKD and other erythroid metabolic genetic disorders.
AB - Pyruvate kinase deficiency (PKD) is a monogenic metabolic disease caused by mutations in the PKLR gene that leads to hemolytic anemia of variable symptomatology and that can be fatal during the neonatal period. PKD recessive inheritance trait and its curative treatment by allogeneic bone marrow transplantation provide an ideal scenario for developing gene therapy approaches. Here, we provide a preclinical gene therapy for PKD based on a lentiviral vector harboring the hPGK eukaryotic promoter that drives the expression of the PKLR cDNA. This therapeutic vector was used to transduce mouse PKD hematopoietic stem cells (HSCs) that were subsequently transplanted into myeloablated PKD mice. Ectopic RPK expression normalized the erythroid compartment correcting the hematological phenotype and reverting organ pathology. Metabolomic studies demonstrated functional correction of the glycolytic pathway in RBCs derived from genetically corrected PKD HSCs, with no metabolic disturbances in leukocytes. The analysis of the lentiviral insertion sites in the genome of transplanted hematopoietic cells demonstrated no evidence of genotoxicity in any of the transplanted animals. Overall, our results underscore the therapeutic potential of the hPGK-coRPK lentiviral vector and provide high expectations toward the gene therapy of PKD and other erythroid metabolic genetic disorders.
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U2 - 10.1038/mt.2016.87
DO - 10.1038/mt.2016.87
M3 - Article
AN - SCOPUS:84982082493
VL - 24
SP - 1187
EP - 1198
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 7
ER -