Adeno-associated virus compositions for restoring HBB gene function and methods of use thereof
Inventors
Chatterjee, Saswati • Wong, Jr., Kamehameha K. • BenHajSalah, Marwa • Smith, Laura Jane • Seymour, Albert Barnes • Wright, Jason Boke • McSwiggen, James Anthony • Dollive, Serena Nicole • St. Martin, Thia Baboval • Prout, Jaime Michelle
Assignees
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Publication Number
US-12201657-B2
Publication Date
2025-01-21
Expiration Date
Abstract
Provided herein are adeno-associated virus (AAV) compositions for correcting a mutation in a beta globin gene (HBB) gene and methods of using the same to correct an HBB gene mutation in a cell. Also provided are packaging systems for making the adeno-associated virus compositions.
Core Innovation
The invention relates to a replication-defective adeno-associated virus (AAV) for editing a target locus in an HBB gene. The AAV includes an AAV capsid comprising an AAV Clade F capsid protein and a correction genome arranged from 5′ to 3′ with a 5′ AAV ITR nucleotide sequence, a 5′ homology arm, an editing element for editing the HBB target locus, a 3′ homology arm, and a 3′ AAV ITR nucleotide sequence.
The correction genome is constrained such that the 5′ homology arm, the editing element, and the 3′ homology arm consist of nucleotide sequence sets identified in the disclosure, including SEQ ID NO: 34 or SEQ ID NO: 38. The disclosed genome architecture uses 5′ and 3′ AAV ITRs with specified nucleotide sequences, SEQ ID NO: 18 for the 5′ AAV ITR and SEQ ID NO: 19 for the 3′ AAV ITR, to provide a homologous recombination-based editing construct centered on the HBB gene.
The disclosure further characterizes AAV-based, nuclease-free in vivo HBB mutation correction using replication-defective AAVs. It is described as correcting HBB for sickle cell disease and beta thalassemia by transducing hematopoietic/erythroid progenitor cells without co-delivering an exogenous nuclease/Cas9.
Claims Coverage
The provided excerpt contains two independent claims. Both independently require a replication-defective AAV with an AAV Clade F capsid and an HBB-targeted correction genome arranged from 5′ to 3′ with defined AAV ITR sequences and constrained homology arm and editing element nucleotide sequences.
Replication-defective AAV with Clade F capsid and SEQ ID NO: 34 correction genome for HBB editing
A replication-defective AAV comprising an AAV capsid comprising an AAV Clade F capsid protein, and a correction genome comprising from 5′ to 3′: a 5′ AAV ITR nucleotide sequence as set forth in SEQ ID NO: 18; a 5′ homology arm nucleotide sequence; an editing element for editing a target locus in an HBB gene; a 3′ homology arm nucleotide sequence; wherein the 5′ homology arm, the editing element and the 3′ homology arm consists of the nucleotide sequence as set forth in SEQ ID NO: 34; and a 3′ AAV ITR nucleotide sequence as set forth in SEQ ID NO: 19.
Replication-defective AAV with Clade F capsid and SEQ ID NO: 38 correction genome for HBB editing
A replication-defective AAV comprising an AAV capsid comprising an AAV Clade F capsid protein, and a correction genome comprising from 5′ to 3′: a 5′ AAV ITR nucleotide sequence as set forth in SEQ ID NO: 18; a 5′ homology arm nucleotide sequence; an editing element for editing a target locus in an HBB gene; a 3′ homology arm nucleotide sequence; wherein the 5′ homology arm, the editing element and the 3′ homology arm consists of the nucleotide sequence as set forth in SEQ ID NO: 38; and a 3′ AAV ITR nucleotide sequence as set forth in SEQ ID NO: 19.
Across the independent claims, the core claim scope is directed to replication-defective AAV vectors using an AAV Clade F capsid and carrying an HBB editing correction genome. The main differentiator between the two provided independent claims is the specific nucleotide sequence constraint for the 5′ homology arm, editing element, and 3′ homology arm, while the AAV ITR sequences are specified as SEQ ID NO: 18 (5′) and SEQ ID NO: 19 (3′).
Stated Advantages
Nuclease-free HBB mutation correction without co-delivering an exogenous nuclease/Cas9.
Documented Applications
Treating sickle cell disease by HBB gene mutation correction using the disclosed AAV approach.
Treating beta thalassemia by HBB gene mutation correction using the disclosed AAV approach.
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