TY - JOUR
T1 - Identification of a myotropic AAV by massively parallel in vivo evaluation of barcoded capsid variants
AU - Weinmann, Jonas
AU - Weis, Sabrina
AU - Sippel, Josefine
AU - Tulalamba, Warut
AU - Remes, Anca
AU - El Andari, Jihad
AU - Herrmann, Anne-Kathrin
AU - Pham, Quang H
AU - Borowski, Christopher
AU - Hille, Susanne
AU - Schönberger, Tanja
AU - Frey, Norbert
AU - Lenter, Martin
AU - VandenDriessche, Thierry
AU - Müller, Oliver J
AU - Chuah, Marinee K
AU - Lamla, Thorsten
AU - Grimm, Dirk
PY - 2020/10/28
Y1 - 2020/10/28
N2 - Adeno-associated virus (AAV) forms the basis for several commercial gene therapy products and for countless gene transfer vectors derived from natural or synthetic viral isolates that are under intense preclinical evaluation. Here, we report a versatile pipeline that enables the direct side-by-side comparison of pre-selected AAV capsids in high-throughput and in the same animal, by combining DNA/RNA barcoding with multiplexed next-generation sequencing. For validation, we create three independent libraries comprising 183 different AAV variants including widely used benchmarks and screened them in all major tissues in adult mice. Thereby, we discover a peptide-displaying AAV9 mutant called AAVMYO that exhibits superior efficiency and specificity in the musculature including skeletal muscle, heart and diaphragm following peripheral delivery, and that holds great potential for muscle gene therapy. Our comprehensive methodology is compatible with any capsids, targets and species, and will thus facilitate and accelerate the stratification of optimal AAV vectors for human gene therapy.
AB - Adeno-associated virus (AAV) forms the basis for several commercial gene therapy products and for countless gene transfer vectors derived from natural or synthetic viral isolates that are under intense preclinical evaluation. Here, we report a versatile pipeline that enables the direct side-by-side comparison of pre-selected AAV capsids in high-throughput and in the same animal, by combining DNA/RNA barcoding with multiplexed next-generation sequencing. For validation, we create three independent libraries comprising 183 different AAV variants including widely used benchmarks and screened them in all major tissues in adult mice. Thereby, we discover a peptide-displaying AAV9 mutant called AAVMYO that exhibits superior efficiency and specificity in the musculature including skeletal muscle, heart and diaphragm following peripheral delivery, and that holds great potential for muscle gene therapy. Our comprehensive methodology is compatible with any capsids, targets and species, and will thus facilitate and accelerate the stratification of optimal AAV vectors for human gene therapy.
KW - myotropic AAV
KW - barcoded capsid variants
KW - in vivo
KW - Adeno-associated virus
KW - gene therapy
KW - gene transfer vectors
KW - DNA/RNA barcoding
U2 - 10.1038/s41467-020-19230-w
DO - 10.1038/s41467-020-19230-w
M3 - Article
C2 - 33116134
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 5432
ER -