Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems

Inventors

Burns, Joseph • ELLIS, Kathryn • Palermo, Adam • SCHWANDER, MARTIN • WHITTON, Jonathon

Assignees

Decibel Therapeutics Inc

Abstract

The disclosure features compositions and methods for the treatment of sensorineural hearing loss and auditory neuropathy, particularly forms of the disease that are associated with mutations in otoferlin (OTOF), by way of OTOF gene therapy. The disclosure provides a variety of compositions that include a first nucleic acid vector that contains a polynucleotide encoding an N-terminal portion of an OTOF protein and a second nucleic acid vector that contains a polynucleotide encoding a C-terminal portion of an OTOF protein. These vectors can be used to increase the expression of OTOF in a subject, such as a human subject suffering from sensorineural hearing loss.

Core Innovation

The invention relates to dual-vector nucleic acid compositions for delivering an otoferlin (OTOF) expression program in which two separate vectors are designed so that neither vector alone encodes a full-length OTOF protein. A first nucleic acid vector includes a myosin 15 (Myo15) promoter operably linked to a first coding polynucleotide encoding an N-terminal portion of an OTOF protein, and a second nucleic acid vector includes a second coding polynucleotide encoding a C-terminal portion of an OTOF protein and a polyadenylation (poly(A)) sequence positioned at a 3′ end.

When introduced into a mammalian cell, the first and second nucleic acid vectors undergo homologous recombination or concatemerization to form a recombined nucleic acid encoding a full-length OTOF protein. The described design rules include overlapping dual vectors in which an overlap is centered at OTOF exon boundaries, with optional inclusion or exclusion of OTOF UTRs, and trans-splicing dual vectors using a splice donor and a splice acceptor positioned relative to non-overlapping N- and C-terminal portions.

The disclosure further describes dual hybrid vectors that combine a recombinogenic region with splice donor and splice acceptor signals, together with regulatory element architectures such as WPRE sequences and vector format options including adeno-associated virus (AAV) vectors with functional ITRs. It also discusses promoter and vector design considerations, including Myo15 promoter sequence-region definitions and the use of rAAV delivery options.

Claims Coverage

The claim coverage centers on a two-vector split-OTOF composition that splits an OTOF coding sequence into N-terminal and C-terminal portions and relies on homologous recombination or concatemerization in a mammalian cell to reconstitute full-length OTOF. Dependent refinements add promoter identity, regulatory elements, splice donor and splice acceptor architecture, exon-boundary splitting, and AAV vector format across 5 inventive features.

The claim coverage centers on two separate nucleic acid vectors that split OTOF into N-terminal and C-terminal portions, with a Myo15 promoter and a 3′ poly(A)-bearing C-terminal construct, and that assemble by homologous recombination or concatemerization in mammalian cells to produce full-length OTOF. Dependent refinements further specify splice donor and splice acceptor plus recombinogenic regions, non-overlapping coding portions, an OTOF exon-boundary split, and AAV vector format.

Stated Advantages

Documented Applications