Methods of treating or preventing pyruvate kinase deficiency

Inventors

SEGOVIA, Jose C. • GOMEZ, Maria G. • NAVARRO, Susana • MEZA, Nestor • BUEREN, Juan • BRAVO, Maria G.

Assignees

Centro de Investigaciones Energeticas Medioambientales y Tecnologicas CIEMAT • Centro de Investigacion Biomedica en Red CIBER • Instituto de Investigacion Sanitaria Fundacion Jimenez Diaz

Abstract

The present disclosure provides polynucleotide cassettes, expression vectors and methods for the expression of a gene in mammalian cells to provide gene therapy for pyruvate kinase deficiency.

Core Innovation

The invention provides a gene-therapy approach for treating or preventing Pyruvate Kinase Deficiency (PKD) in a subject in need thereof. The approach involves contacting a hematopoietic stem cell with a recombinant lentivirus vector and administering the hematopoietic stem cell to the subject. The recombinant lentivirus vector comprises an expression cassette encoding a pyruvate kinase, liver and red blood cell (PKLR) polypeptide operatively linked to a promoter.

The expression cassette further includes a woodchuck hepatitis virus post-transcriptional regulatory element (Wpre). The Wpre comprises a sequence at least 90% identical to SEQ ID NO:1, including chimeric/mutated Wpre sequence variants and sequence identity variants.

The partial content also describes preclinical/translation context for PKD, including correction of anemia and reticulocytosis in PKD mice following PGK-coRPK lentiviral transduction and transplantation of lineage-depleted bone marrow/hematopoietic stem cells, along with normalization of erythroid differentiation and rescue of spleen/liver pathology and iron overload. It further describes genome-wide lentiviral vector integration mapping showing polyclonal reconstitution and lack of common insertion sites, with genome-wide integration analysis not enriching for oncogenic pathways based on Gene Ontology (GO) analysis.

Claims Coverage

The partial content provides two independent claims. Each independent claim is focused on the same core inventive architecture: a recombinant lentivirus vector with an expression cassette encoding PKLR, operatively linked to a promoter, and including a Wpre sequence with high sequence identity to SEQ ID NO:1. Across the two independent claims, the inventive features are captured by the lentiviral expression cassette design and the use of hematopoietic stem cells for treatment of PKD.

Across the independent claims, the inventive claim coverage is directed to PKD treatment methods that use hematopoietic stem cells modified with a recombinant lentivirus vector encoding PKLR under a promoter with a woodchuck hepatitis virus Wpre sequence having at least 90% identity to SEQ ID NO:1. One claim requires contacting then administering the hematopoietic stem cell, and the other requires administering the subject transduced hematopoietic stem cell.

Stated Advantages

Documented Applications