Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells

B. Cheng, C. Ling, Y. Dai, Y. Lu, L. G. Glushakova, S. W.Y. Gee, K. E. McGoogan, G. V. Aslanidi, M. Park, P. W. Stacpoole, D. Siemann, Chen Liu, A. Srivastava, C. Ling

Research output: Contribution to journalArticle

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Abstract

Our recent studies have revealed that among the 10 different commonly used adeno-associated virus (AAV) serotypes, AAV3 vectors transduce human liver cancer cells extremely efficiently because these cells express high levels of human hepatocyte growth factor receptor (hHGFR), and AAV3 utilizes hHGFR as a cellular co-receptor for viral entry. In this report, we provide further evidence that both extracellular as well as intracellular kinase domains of hHGFR are involved in AAV3 vector entry and AAV3-mediated transgene expression. We also document that AAV3 vectors are targeted for degradation by the host cell proteasome machinery, and that site-directed mutagenesis of surface-exposed tyrosine (Y) to phenylalanine (F) residues on AAV3 capsids significantly improves the transduction efficiency of Y701F, Y705F and Y731F mutant AAV3 vectors. The transduction efficiency of the Y705+731F double-mutant vector is significantly higher than each of the single mutants in liver cancer cells in vitro. In immunodeficient mouse xenograft models, direct intratumoral injection of AAV3 vectors also led to high-efficiency transduction of human liver tumor cells in vivo. We also document here that the optimized tyrosine-mutant AAV3 vectors lead to increased transduction efficiency following both intratumoral and tail-vein injections in vivo. The optimized tyrosine-mutant AAV3 serotype vectors containing proapoptotic genes should prove useful for the potential gene therapy of human liver cancers.

Original languageEnglish (US)
Pages (from-to)375-384
Number of pages10
JournalGene therapy
Volume19
Issue number4
DOIs
StatePublished - Apr 1 2012
Externally publishedYes

Fingerprint

Liver Neoplasms
Proto-Oncogene Proteins c-met
Tyrosine
Dependovirus
Injections
Capsid
Proteasome Endopeptidase Complex
Site-Directed Mutagenesis
Phenylalanine
Transgenes
Heterografts
Genetic Therapy
Tail
Veins
Phosphotransferases
Serogroup
Liver
Genes
human HGF protein
Neoplasms

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Cheng, B., Ling, C., Dai, Y., Lu, Y., Glushakova, L. G., Gee, S. W. Y., ... Ling, C. (2012). Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells. Gene therapy, 19(4), 375-384. https://doi.org/10.1038/gt.2011.105
Cheng, B. ; Ling, C. ; Dai, Y. ; Lu, Y. ; Glushakova, L. G. ; Gee, S. W.Y. ; McGoogan, K. E. ; Aslanidi, G. V. ; Park, M. ; Stacpoole, P. W. ; Siemann, D. ; Liu, Chen ; Srivastava, A. ; Ling, C. / Development of optimized AAV3 serotype vectors : Mechanism of high-efficiency transduction of human liver cancer cells. In: Gene therapy. 2012 ; Vol. 19, No. 4. pp. 375-384.
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Cheng, B, Ling, C, Dai, Y, Lu, Y, Glushakova, LG, Gee, SWY, McGoogan, KE, Aslanidi, GV, Park, M, Stacpoole, PW, Siemann, D, Liu, C, Srivastava, A & Ling, C 2012, 'Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells', Gene therapy, vol. 19, no. 4, pp. 375-384. https://doi.org/10.1038/gt.2011.105

Development of optimized AAV3 serotype vectors : Mechanism of high-efficiency transduction of human liver cancer cells. / Cheng, B.; Ling, C.; Dai, Y.; Lu, Y.; Glushakova, L. G.; Gee, S. W.Y.; McGoogan, K. E.; Aslanidi, G. V.; Park, M.; Stacpoole, P. W.; Siemann, D.; Liu, Chen; Srivastava, A.; Ling, C.

In: Gene therapy, Vol. 19, No. 4, 01.04.2012, p. 375-384.

Research output: Contribution to journalArticle

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T1 - Development of optimized AAV3 serotype vectors

T2 - Mechanism of high-efficiency transduction of human liver cancer cells

AU - Cheng, B.

AU - Ling, C.

AU - Dai, Y.

AU - Lu, Y.

AU - Glushakova, L. G.

AU - Gee, S. W.Y.

AU - McGoogan, K. E.

AU - Aslanidi, G. V.

AU - Park, M.

AU - Stacpoole, P. W.

AU - Siemann, D.

AU - Liu, Chen

AU - Srivastava, A.

AU - Ling, C.

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