ENPP1 gene therapy for the treatment of vascular disease
AssigneesThe General Hospital Corporation, The Brigham and Women's Hospital, Inc.
InventorsRajeev Malhotra, Patricia L. Musolino, Mark Evan Lindsay, Natalie Artzi, Eliz Amar-Lewis, Christian Lacks Lino Cardenas
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Abstract
Provided herein are compositions and methods for gene therapy for disorders of arterial calcification as well as Generalized Arterial Calcification of Infancy (GACI). The methods include a gene addition strategy to deliver a DNA construct to target tissues (such as liver and smooth muscle cells) to express soluble recombinant ENPP1 (srENPP1) or transmembrane full-length recombinant ENPP1 (rENPP1).
Claims20
A recombinant adeno associated viral vector (AAV) encapsulating a construct encoding an ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) enzyme, said vector comprising:
- an expression cassette comprising:
- a 5′ ITR;
- a promoter;
- a nucleic acid encoding an ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) enzyme, wherein said ENPP1 enzyme is a full-length transmembrane ENPP1 enzyme that comprises SEQ ID NO:1 or comprises a sequence that is at least 95% identical to SEQ ID NO:1;
- and
- a 3′ ITR;
- and
- wherein the recombinant AAV vector comprises a capsid with a nucleic encoding peptide sequence PRPPSTH (SEQ ID NO:44) inserted therein.
A nucleic acid construct for expression of an ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) enzyme, said nucleic acid construct comprising:
- a nucleic acid sequence encoding an AAV genome comprising:
- a first inverted terminal repeats (ITR);
- a nucleic acid encoding a replication (rep) sequence;
- a nucleic acid encoding a capsid (cap) sequence, the nucleic acid encoding the cap sequence comprising a sequence encoding the peptide PRPPSTH inserted therein (SEQ ID NO:44);
- a promoter;
- a nucleic acid encoding an ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) enzyme comprising a transmembrane domain, wherein said ENPP1 enzyme is a full-length transmembrane ENPP1 enzyme comprising SEQ ID NO:1 or comprises a sequence that is at least 95% identical to SEQ ID NO:1;
- and
- a second ITR.
A particle comprising:
- a) an amount of an ionizable lipid;
- an amount of neutral lipid;
- an amount of cholesterol;
- an amount of one or more PEG-lipids;
- and
- an amount of a DOTAP molecule;
- b) a peptide conjugated to a linker in the particle;
- and
- c) a construct for expression of a full-length transmembrane ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), comprising a sequence encoding a full-length ENPP1 transgene, wherein the ENPP1 is (i) full-length human transmembrane ENPP1, and a promoter that drives expression of the ENPP1 transgene.
Dependencies are inferred from the claim text.