Gene therapies for lysosomal disorders
Inventors
Abeliovich, Asa • Heckman, Laura • RHINN, Herve
Assignees
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Publication Number
US-11807849-B2
Publication Date
2023-11-07
Expiration Date
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 one or more inhibitory nucleic acids targeting SCNA or a portion thereof, TMEM106B or a portion thereof, 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 concepts for lysosomal/PD-related disorders, using expression constructs that deliver inhibitory nucleic acids targeting α-Synuclein. The inhibitory nucleic acid is encoded by the sequence set forth in SEQ ID NO: 3 and is provided as a transgene encoding a single gene product. The nucleic acid is configured as an expression construct flanked by two adeno-associated virus (AAV) inverted terminal repeats (ITR) sequences, including a 5′ ITR and a 3′ ITR.
The invention further describes recombinant AAV vector architectures that include regulatory and processing elements associated with the expression construct. A representative rAAV vector genome organization includes a CMV enhancer, a CBA promoter, WPRE, and a Bovine Growth Hormone polyA signal tail, positioned between the 5′ AAV ITR and the 3′ AAV ITR. The disclosed designs also include specific arrangement constraints using defined nucleotide intervals and defined ITR sequence content, including specified ITR “D” region concepts.
The disclosed subject matter additionally covers AAV ITR and genome configurations involving modified ITR “D” regions, including positioning concepts where the modified “D” region is located outside vs wild-type “inside” relative to the expression construct. The document also includes rAAV capsid protein options capable of BBB crossing, and describes treatment-related implementations for Parkinson’s disease via CNS or peripheral administration, along with example systems used to evaluate the inhibitory nucleic acid targeting α-Synuclein.
Claims Coverage
The provided independent claims cover multiple categories of protected subject matter—isolated nucleic acids, rAAV vectors, rAAV genome constructions with defined regulatory elements and order, baculovirus vectors, and isolated cells with a rep/cap system—centered on delivering a single inhibitory nucleic acid targeting α-Synuclein encoded by SEQ ID NO: 3 using AAV ITR-flanked expression constructs. Across the independent claims, the inventive features include the specific inhibitory-nucleic-acid sequence, the AAV ITR-flanked expression construct, and multiple defined vector/cell implementations and genome element arrangements.
AAV ITR-flanked inhibitory nucleic acid targeting α-Synuclein in an isolated nucleic acid
An isolated nucleic acid comprising (i) an expression construct comprising a transgene encoding a single gene product, wherein the gene product is an inhibitory nucleic acid targeting α-Synuclein, wherein the inhibitory nucleic acid is encoded by the sequence set forth in SEQ ID NO: 3, and (ii) two adeno-associated virus (AAV) inverted terminal repeats (ITR) sequences flanking the expression construct, wherein the first ITR sequence is a 5′ ITR, and the second ITR sequence is a 3′ ITR.
An rAAV vector with an inhibitory nucleic acid targeting α-Synuclein flanked by 5′ and 3′ AAV ITR
A recombinant adeno-associated virus (rAAV) vector comprising (i) an expression construct comprising a transgene encoding a single gene product, wherein the gene product is an inhibitory nucleic acid targeting α-Synuclein, wherein the inhibitory nucleic acid is encoded by the sequence set forth in SEQ ID NO: 3, and (ii) two AAV ITR sequences flanking the expression construct, wherein the first ITR sequence is a 5′ ITR, and the second ITR sequence is a 3′ ITR.
Order-defined rAAV genome including CMV enhancer, CBA promoter, WPRE, polyA tail, and ITRs
A rAAV vector comprising a nucleic acid comprising, in 5′ to 3′ order: (a) a 5′ AAV ITR; (b) a CMV enhancer; (c) a CBA promoter; (d) a transgene encoding a single gene product, wherein the gene product is an inhibitory nucleic acid targeting α-Synuclein, wherein the inhibitory nucleic acid is encoded by the sequence set forth in SEQ ID NO: 3; (e) a WPRE; (f) a Bovine Growth Hormone polyA signal tail; and (g) a 3′ AAV ITR.
Baculovirus vector carrying SEQ ID NO: 3 inhibitory nucleic acid flanked by AAV ITRs
A Baculovirus vector comprising a transgene encoding a single gene product, wherein the gene product is an inhibitory nucleic acid coding sequence comprising the sequence set forth in SEQ ID NO: 3, and two AAV ITR sequences flanking the sequence set forth in SEQ ID NO: 3.
Isolated cell with separate rep/cap vector(s) and a second AAV ITR-flanked inhibitory nucleic acid vector
An isolated cell comprising: (i) a first vector encoding one or more AAV rep proteins and/or one or more AAV cap proteins; and (ii) a second vector comprising a transgene encoding a single gene product, wherein the gene product is an inhibitory nucleic acid coding sequence comprising the sequence set forth in SEQ ID NO: 3, and two AAV ITR sequences flanking the sequence set forth in SEQ ID NO: 3.
Collectively, the independent claims provide coverage for nucleic-acid and vector systems (including rAAV and baculovirus), and for isolated cells containing a rep/cap delivery system plus an AAV ITR-flanked expression construct, where the expression construct contains an inhibitory nucleic acid targeting α-Synuclein encoded by SEQ ID NO: 3.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Parkinson’s disease (PD) treatment via CNS or peripheral administration.
Gaucher disease context within lysosomal/PD-related disorders.
Cell assays including in vitro knockdown validation for α-Synuclein (SCNA) and TMEM106B.
Mutant mice and chemically induced Gaucher models.
Clinical trials in PD/LBD/Gaucher disease.
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