Gene therapies for lysosomal disorders

Inventors

Abeliovich, Asa • Heckman, Laura • RHINN, Herve

Assignees

Prevail Therapeutics Inc

Abstract

The disclosure relates, in some aspects, to compositions and methods for treatment of diseases associated with aberrant lysosomal function, for example Parkinson's disease and Gaucher disease. In some embodiments, the disclosure provides expression constructs comprising a transgene encoding beta-Glucocerebrosidase (GBA) or a portion thereof, Lysosomal Membrane Protein 2 (LIMP2), Prosaposin, or any combination of the foregoing. In some embodiments, the disclosure provides methods of Parkinson's disease by administering such expression constructs to a subject in need thereof.

Core Innovation

The disclosed invention relates to gene-therapy compositions and methods for lysosomal disorders, including Gaucher disease, and for Parkinson’s disease associated with GBA1-related mechanisms. The approach uses AAV-based expression constructs encoding a lysosomal Gcase protein, with the transgene insert encoding a Gcase protein comprising the nucleotide sequence of SEQ ID NO: 15, and with an expression construct flanked by two AAV inverted terminal repeat (ITR) sequences.

The constructs are further defined by the arrangement and selection of regulatory elements, including a promoter operably linked to the transgene insert, and optionally including enhancers and RNA regulatory elements. In representative configurations, the nucleic acid includes a CMVe enhancer, a CBA promoter, WPRE, and a bovine growth hormone polyA signal tail, with specific ordering in 5′ to 3′ order together with 5′ and 3′ AAV ITR sequences.

A key design aspect described in the document is the modified AAV ITR “D” region placement/sequence compared to wild-type AAV ITRs, including references to a wild-type AAV2 ITR (SEQ ID NO: 29) and a modified “D” region (SEQ ID NOs: 26 and 27). The document also describes inclusion of a TRY region (SEQ ID NO: 28) positioned between the 5′ ITR and the expression construct in certain construct architectures, and relates these construct features to nonclinical characterization in lysosomal and Parkinson-relevant models.

Claims Coverage

The independent claims in the provided set cover nucleic acid expression constructs and associated delivery/production components for AAV-mediated expression of a Gcase (beta-glucocerebrosidase) transgene defined by SEQ ID NO: 15. The inventive features are expressed through constraints on the transgene sequence, the promoter-transgene operable linkage, and the specific flanking AAV ITR arrangement, with additional independent claims directed to plasmid, baculovirus, isolated cells/vector-production systems, and rAAV vectors.

Overall, the claim coverage centers on AAV ITR-flanked expression constructs encoding a Gcase transgene defined by SEQ ID NO: 15, with independent claims spanning isolated nucleic acids with defined ITR and regulatory element content/order, and extending to plasmid, baculovirus vector, two-vector isolated cell systems for AAV packaging, and rAAV vectors carrying the SEQ ID NO: 15 Gcase expression construct.

Stated Advantages

Documented Applications