Directed evolution of a family of AAV capsid variants enabling potent muscle-directed gene delivery across species

Mohammadsharif Tabebordbar; Kim A Lagerborg; Alexandra Stanton; Emily M King; Simon Ye; Liana Tellez; Allison Krunnfusz; Sahar Tavakoli; Jeffrey J Widrick; Kathleen A Messemer; Emily C Troiano; Behzad Moghadaszadeh; Bryan L Peacker; Krystynne A Leacock; Naftali Horwitz; Alan H Beggs; Amy J Wagers; Pardis C Sabeti

doi:10.1016/j.cell.2021.08.028

Directed evolution of a family of AAV capsid variants enabling potent muscle-directed gene delivery across species

Cell. 2021 Sep 16;184(19):4919-4938.e22. doi: 10.1016/j.cell.2021.08.028. Epub 2021 Sep 9.

Authors

Mohammadsharif Tabebordbar¹, Kim A Lagerborg², Alexandra Stanton³, Emily M King⁴, Simon Ye⁵, Liana Tellez⁴, Allison Krunnfusz⁴, Sahar Tavakoli⁶, Jeffrey J Widrick⁷, Kathleen A Messemer⁸, Emily C Troiano⁷, Behzad Moghadaszadeh⁷, Bryan L Peacker⁸, Krystynne A Leacock⁸, Naftali Horwitz⁹, Alan H Beggs¹⁰, Amy J Wagers¹¹, Pardis C Sabeti¹²

Affiliations

¹ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: [email protected].
² Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, MA 02115, USA.
³ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard Program in Virology, Harvard Medical School, Boston, MA 02115, USA.
⁴ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
⁵ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard-MIT Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
⁶ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA; Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital, Boston, MA 02115, USA.
⁷ Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
⁸ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA.
⁹ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA; Section on Islet Cell and Regenerative Biology, Joslin Diabetes Center, Boston, MA 02215, USA.
¹⁰ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
¹¹ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA; Section on Islet Cell and Regenerative Biology, Joslin Diabetes Center, Boston, MA 02215, USA. Electronic address: [email protected].
¹² Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Organismic and Evolutionary Biology, FAS Center for Systems Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA. Electronic address: [email protected].

Abstract

Replacing or editing disease-causing mutations holds great promise for treating many human diseases. Yet, delivering therapeutic genetic modifiers to specific cells in vivo has been challenging, particularly in large, anatomically distributed tissues such as skeletal muscle. Here, we establish an in vivo strategy to evolve and stringently select capsid variants of adeno-associated viruses (AAVs) that enable potent delivery to desired tissues. Using this method, we identify a class of RGD motif-containing capsids that transduces muscle with superior efficiency and selectivity after intravenous injection in mice and non-human primates. We demonstrate substantially enhanced potency and therapeutic efficacy of these engineered vectors compared to naturally occurring AAV capsids in two mouse models of genetic muscle disease. The top capsid variants from our selection approach show conserved potency for delivery across a variety of inbred mouse strains, and in cynomolgus macaques and human primary myotubes, with transduction dependent on target cell expressed integrin heterodimers.

Keywords: AAV capsid engineering; Duchenne muscular dystrophy; MyoAAV; X-linked myotubular myopathy; directed evolution; integrin heterodimers; muscle gene therapy; non-human primates.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Capsid / chemistry
Capsid / metabolism*
Cells, Cultured
Dependovirus / metabolism*
Directed Molecular Evolution*
Disease Models, Animal
Gene Transfer Techniques*
HEK293 Cells
Humans
Integrins / metabolism
Macaca fascicularis
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Muscle Fibers, Skeletal / metabolism
Muscle, Skeletal / metabolism*
Muscular Dystrophy, Duchenne / pathology
Muscular Dystrophy, Duchenne / therapy
Myopathies, Structural, Congenital / pathology
Myopathies, Structural, Congenital / therapy
Protein Multimerization
Protein Tyrosine Phosphatases, Non-Receptor / genetics
Protein Tyrosine Phosphatases, Non-Receptor / metabolism
Protein Tyrosine Phosphatases, Non-Receptor / therapeutic use
Recombination, Genetic / genetics
Species Specificity
Transgenes

Substances

Integrins
Protein Tyrosine Phosphatases, Non-Receptor
myotubularin

Abstract

Publication types

MeSH terms

Substances

Grants and funding