Powers and colleagues have developed an AAV MS-based multi-attribute method (MAM) to quantitate deamidation, an important quality attribute for AAV therapeutics. The method was validated according to ...

Kontogiannis and colleagues developed a mass spectrometry-based method for quantification of the capsid proteins of recombinant AAV9 and measurement of the mean capsid stoichiometry.

Kontogiannis and colleagues developed a mass spectrometry-based method for quantification of the capsid proteins of recombinant AAV9 and measurement of the mean capsid stoichiometry.

This paper details development and use of a sensitive, cell-based assay for detection of replication-competent AAV (rcAAV) in AAV viral vector preparations, suitable for quality control testing of clinical drug substances for gene therapy applications.

AAV8-mediated gene therapy targeting the liver effectively normalized peripheral oxysterol levels in a mouse model of hereditary spastic paraplegia type 5 (SPG5). However, brain oxysterol levels remained partially corrected, indicating that successful SPG5 treatment requires a gene therapy approach that also targets the central nervous system.

Designed chimeric PCSK9 vaccine elicits an adequate humoral immune response without causing systemic inflammation or destruction of healthy tissue. Vaccine efficacy is demonstrated by reduced serum cholesterol and suppresed atherosclerosis development in the aortic tissue. This represents a proof-of-concept for vaccines targeting endogenous proteins.

Powers and colleagues have developed an AAV MS-based multi-attribute method (MAM) to quantitate deamidation, an important quality attribute for AAV therapeutics. The method was validated according to according to ICH Q2(R2) and implemented for routine testing of AAV for development and stability purposes.

Ling and colleagues identified transgene overexpression-induced cytotoxicity in their previously reported AAV9- based gene replacement therapy for SURF1- related Leigh syndrome. By optimizing the vector design, they now developed an improved version that showed efficacy equivalent to their original design, but without associated cytotoxicity in wildtype rats.

This study introduces and validates Quantitative Electron Microscopy (QuTEM) as a robust method for characterizing adeno-associated virus (AAV) vectors by accurately distinguishing full, partial, and empty capsids. QuTEM's reliability enhances quality control in AAV production, ensuring therapeutic efficacy in gene therapy applications. This advancement is crucial for optimizing AAV vector characterization.

Ma and colleagues identified superior blood-brain barrier (BBB)-crossing AAV variants effective in both C57BL/6J and BALB/c mice through a capsid screen and systematically evaluated the potential of eight different capsid engineering strategies for discovering novel variants.

Microdystrophins are miniaturized versions of dystrophin, used in AAV gene therapy of Duchenne muscular dystrophy. Danos and colleagues describe an enhanced microdystrophin with a C-terminal extension that better protects dystrophic muscle against contraction-induced damage and improves recovery from injury relative to an equivalent protein lacking the C-terminal domain.

Boink and colleagues demonstrate that AAV6 provides superior transduction efficiency over AAV9 and engineered MyoAAV4A in locally applied gene therapy in vitro, ex vivo and in vivo in various rodent, pig and human models. These findings underscore the robust transduction efficiency with AAV6 in the setting of direct intramyocardial injection.

Plückthun and colleagues present a novel genome-replicating high-capacity adenoviral vector that enhances in situ transgene expression without inducing cytotoxicity. This new adenoviral vector class combines large payload capacity (≥ 22 kb) with self-induced and self-sustained genome replication, offering a promising tool for improved gene therapies and vaccination strategies.

For clinical relevance, AAV-delivered gene therapies must be contained within single vectors. Jones and colleagues have engineered all-in-one AAV vectors for CRISPR inhibition of pathogenic DUX4 expression in Facioscapulohumeral muscular dystrophy (FSHD). These vectors have been validated in FSHD myocytes and in a human xenograft model of the disease

Rose and colleagues sequenced the transgene of commercially available AAV products using three different sequencing methods. Two methods showed a similar overall per-base mutation rate (consistent across suppliers and production lots) that was higher in the vectors than in the plasmids. However, there was little consistency among methods for identifying the specific location in the gene where the mutations occurred.

Currier and colleagues describe development of a clinical lead adeno-associated virus construct, VNX-101, with enhanced features to achieve off-the-shelf, long-term in vivo T cell engagement for CD19+ B cell malignancies. A single dose of VNX-101 was safe in mice and NHPs and eliminated B cell malignancies in xenograft models.