title: Safe and Efficient Gene Therapy for Pyruvate Kinase Deficiency.
abstract: 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.
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Garcia-Gomez
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Calabria
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Garcia-Bravo
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Benedicenti
authors.affiliation: Agios Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Penelope
authors.initials: P
authors.lastname: Kosinski
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: López-Manzaneda
authors.affiliation: Agios Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Collin
authors.initials: C
authors.lastname: Hill
authors.affiliation: Red Cell Pathology Laboratory. Hospital Clínic of Barcelona - Institut d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Del Mar Mañu-Pereira
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Martín
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Israel
authors.initials: I
authors.lastname: Orman
authors.affiliation: Red Cell Pathology Laboratory. Hospital Clínic of Barcelona - Institut d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain.
authors.forename: Joan-LLuis
authors.initials: JL
authors.lastname: Vives-Corrons
authors.affiliation: Agios Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Kung
authors.affiliation: Institute of Experimental Hematology at Hannover Medical Hospital, Hannover, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Schambach
authors.affiliation: Agios Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Shengfang
authors.initials: S
authors.lastname: Jin
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Bueren
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Montini
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Susana
authors.initials: S
authors.lastname: Navarro
authors.affiliation: Hematopoietic Innovative Therapies Division, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) - Centro de Investigaciones Biomédicas en Red de Enfermedades Raras (CIBERER), Madrid, Spain.|Advanced Therapies Mixed Unit. Instituto de Investigación Sanitaria-Fundación Jiménez Díaz (IIS-FJD), Madrid, Spain.
authors.forename: Jose C
authors.initials: JC
authors.lastname: Segovia
mesh_terms: D000746:Anemia, Hemolytic, Congenital Nonspherocytic / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D000818:Animals; D001773:Blood Cells / Q000378:metabolism; D002454:Cell Differentiation; D004195:Disease Models, Animal; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D004920:Erythropoiesis; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006019:Glycolysis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D055432:Metabolomics; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D010641:Phenotype; D011770:Pyruvate Kinase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D015323:Pyruvate Metabolism, Inborn Errors / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D014161:Transduction, Genetic
chemical_list: D011770:Pyruvate Kinase