Next generation of adeno-associated virus 2 vectors: Point mutations in tyrosines lead to high-efficiency transduction at lower doses

• DOI: 10.1073/pnas.0802866105
• Publication Date: 2008-05-30T00:54:44Z
• Authors: Li Zhong; Baozheng Li; Cathryn S. Mah; Lakshmanan Govind...; Mavis Agbandje-Mc...; Mario Cooper; Roland W. Herzog; Irene Zolotukhin; Kenneth H. Warrin...; Kirsten A. Weigel...; Jacqueline A. Hobbs; Sergei Zolotukhin; Nicholas Muzyczka; Arun Srivastava
• Keywords: Transduction (biophysics)

Recombinant adeno-associated virus 2 (AAV2) vectors are in use several Phase I/II clinical trials, but relatively large vector doses needed to achieve therapeutic benefits. Large also trigger an immune response as a significant fraction of the fails traffic efficiently nucleus and is targeted for degradation by host cell proteasome machinery. We have reported that epidermal growth factor receptor protein tyrosine kinase (EGFR-PTK) signaling negatively affects transduction AAV2 impairing nuclear transport vectors. observed EGFR-PTK can phosphorylate capsids at residues. Tyrosine-phosphorylated enter cells fail transduce effectively, part because ubiquitination AAV followed proteasome-mediated degradation. reasoned mutations surface-exposed residues might allow evade phosphorylation subsequent and, thus, prevent Here, we document site-directed mutagenesis leads production HeLa ≈10-fold more vitro murine hepatocytes nearly 30-fold vivo log lower dose. Therapeutic levels human Factor IX (F.IX) produced reduced The increased efficiency tyrosine-mutant due lack capsid improved intracellular trafficking nucleus. These studies led development capable high-efficiency doses, which has important implications their gene therapy.