Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells

L Zhang; A Rossi; L Lange; N Meumann; U Koitzsch; Kathleen Christie; M. Andrew Nesbit; Tara C. B. Moore; U.T. Hacker; M. Morgan; D. Hoffmann; James Zengel; Jan E. Carette; A. Schambach; A. Salvetti; Margarete Odenthal; Hildegard Büning

doi:10.1089/hum.2019.027

Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells

L Zhang, A Rossi, L Lange, N Meumann, U Koitzsch, Kathleen Christie, M. Andrew Nesbit, Tara C. B. Moore, U.T. Hacker, M. Morgan, D. Hoffmann, James Zengel, Jan E. Carette, A. Schambach, A. Salvetti, Margarete Odenthal, Hildegard Büning

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Abstract

Endothelial cells (EC) are targets in gene therapy and regenerative medicine, but they are inefficiently transduced with adeno-associated virus (AAV) vectors of various serotypes. To identify barriers hampering efficient transduction and to develop an optimized AAV variant for EC transduction, we screened an AAV serotype 2-based peptide display library on primary human macrovascular EC. Using a new high-throughput selection and monitoring protocol, we identified a capsid variant, AAV-V _EC, which outperformed the parental serotype as well as first-generation targeting vectors in EC transduction. AAV vector uptake was improved, resulting in significantly higher transgene expression levels from single-stranded vector genomes detectable within a few hours post-transduction. Notably, AAV-V _EC transduced not only proliferating EC but also quiescent EC, although higher particle-per-cell ratios had to be applied. Also, induced pluripotent stem cell-derived endothelial progenitor cells, a novel tool in regenerative medicine and gene therapy, were highly susceptible toward AAV-V _EC transduction. Thus, overcoming barriers by capsid engineering significantly expands the AAV tool kit for a wide range of applications targeting EC.

Original language	English
Pages (from-to)	1284-1296
Number of pages	13
Journal	Human Gene Therapy
Volume	30
Issue number	10
Early online date	13 Aug 2019
DOIs	https://doi.org/10.1089/hum.2019.027
Publication status	Published (in print/issue) - 23 Sept 2019

Bibliographical note

Funding Information:
This work was supported by grants from University Hospital Cologne (Kln Fortune) to H.B. and M.O., the Center for Molecular Medicine Cologne (CMMC) to H.B., BMBF and MWK Lower Saxony-funded Professorinnenprogramm Niedersachsen to H.B., the DFG-funded KFO286 to H.B., the DFG-funded cluster of excellence REBIRTH EXC62/2 to A.S. and H.B., and the DFG-funded SFB738 to A.S. L.Z. held a scholarship from the German Academic Exchange Service (DAAD).

Publisher Copyright:
© 2019, Mary Ann Liebert, Inc.

Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.

Keywords

AAV vectors
endothelial cells
induced pluripotent stem cells
uncoating
uptake

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1089/hum.2019.027

HUM-2019-027-Zhang_1PAuthor Accepted Manuscript, 1.06 MB

Cite this

Zhang, L., Rossi, A., Lange, L., Meumann, N., Koitzsch, U., Christie, K., Nesbit, M. A., Moore, T. C. B., Hacker, U. T., Morgan, M., Hoffmann, D., Zengel, J., Carette, J. E., Schambach, A., Salvetti, A., Odenthal, M., & Büning, H. (2019). Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells. Human Gene Therapy, 30(10), 1284-1296. https://doi.org/10.1089/hum.2019.027

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title = "Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells",

abstract = "Endothelial cells (EC) are targets in gene therapy and regenerative medicine, but they are inefficiently transduced with adeno-associated virus (AAV) vectors of various serotypes. To identify barriers hampering efficient transduction and to develop an optimized AAV variant for EC transduction, we screened an AAV serotype 2-based peptide display library on primary human macrovascular EC. Using a new high-throughput selection and monitoring protocol, we identified a capsid variant, AAV-V EC, which outperformed the parental serotype as well as first-generation targeting vectors in EC transduction. AAV vector uptake was improved, resulting in significantly higher transgene expression levels from single-stranded vector genomes detectable within a few hours post-transduction. Notably, AAV-V EC transduced not only proliferating EC but also quiescent EC, although higher particle-per-cell ratios had to be applied. Also, induced pluripotent stem cell-derived endothelial progenitor cells, a novel tool in regenerative medicine and gene therapy, were highly susceptible toward AAV-V EC transduction. Thus, overcoming barriers by capsid engineering significantly expands the AAV tool kit for a wide range of applications targeting EC. ",

keywords = "AAV vectors, endothelial cells, induced pluripotent stem cells, uncoating, uptake",

author = "L Zhang and A Rossi and L Lange and N Meumann and U Koitzsch and Kathleen Christie and Nesbit, {M. Andrew} and Moore, {Tara C. B.} and U.T. Hacker and M. Morgan and D. Hoffmann and James Zengel and Carette, {Jan E.} and A. Schambach and A. Salvetti and Margarete Odenthal and Hildegard B{\"u}ning",

note = "Funding Information: This work was supported by grants from University Hospital Cologne (Kln Fortune) to H.B. and M.O., the Center for Molecular Medicine Cologne (CMMC) to H.B., BMBF and MWK Lower Saxony-funded Professorinnenprogramm Niedersachsen to H.B., the DFG-funded KFO286 to H.B., the DFG-funded cluster of excellence REBIRTH EXC62/2 to A.S. and H.B., and the DFG-funded SFB738 to A.S. L.Z. held a scholarship from the German Academic Exchange Service (DAAD). Publisher Copyright: {\textcopyright} 2019, Mary Ann Liebert, Inc. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.",

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doi = "10.1089/hum.2019.027",

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Zhang, L, Rossi, A, Lange, L, Meumann, N, Koitzsch, U, Christie, K, Nesbit, MA , Moore, TCB, Hacker, UT, Morgan, M, Hoffmann, D, Zengel, J, Carette, JE, Schambach, A, Salvetti, A, Odenthal, M & Büning, H 2019, 'Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells', Human Gene Therapy, vol. 30, no. 10, pp. 1284-1296. https://doi.org/10.1089/hum.2019.027

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AU - Zhang, L

AU - Rossi, A

AU - Lange, L

AU - Meumann, N

AU - Koitzsch, U

AU - Christie, Kathleen

AU - Nesbit, M. Andrew

AU - Moore, Tara C. B.

AU - Hacker, U.T.

AU - Morgan, M.

AU - Hoffmann, D.

AU - Zengel, James

AU - Carette, Jan E.

AU - Schambach, A.

AU - Salvetti, A.

AU - Odenthal, Margarete

AU - Büning, Hildegard

N1 - Funding Information: This work was supported by grants from University Hospital Cologne (Kln Fortune) to H.B. and M.O., the Center for Molecular Medicine Cologne (CMMC) to H.B., BMBF and MWK Lower Saxony-funded Professorinnenprogramm Niedersachsen to H.B., the DFG-funded KFO286 to H.B., the DFG-funded cluster of excellence REBIRTH EXC62/2 to A.S. and H.B., and the DFG-funded SFB738 to A.S. L.Z. held a scholarship from the German Academic Exchange Service (DAAD). Publisher Copyright: © 2019, Mary Ann Liebert, Inc. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.

PY - 2019/9/23

Y1 - 2019/9/23

N2 - Endothelial cells (EC) are targets in gene therapy and regenerative medicine, but they are inefficiently transduced with adeno-associated virus (AAV) vectors of various serotypes. To identify barriers hampering efficient transduction and to develop an optimized AAV variant for EC transduction, we screened an AAV serotype 2-based peptide display library on primary human macrovascular EC. Using a new high-throughput selection and monitoring protocol, we identified a capsid variant, AAV-V EC, which outperformed the parental serotype as well as first-generation targeting vectors in EC transduction. AAV vector uptake was improved, resulting in significantly higher transgene expression levels from single-stranded vector genomes detectable within a few hours post-transduction. Notably, AAV-V EC transduced not only proliferating EC but also quiescent EC, although higher particle-per-cell ratios had to be applied. Also, induced pluripotent stem cell-derived endothelial progenitor cells, a novel tool in regenerative medicine and gene therapy, were highly susceptible toward AAV-V EC transduction. Thus, overcoming barriers by capsid engineering significantly expands the AAV tool kit for a wide range of applications targeting EC.

AB - Endothelial cells (EC) are targets in gene therapy and regenerative medicine, but they are inefficiently transduced with adeno-associated virus (AAV) vectors of various serotypes. To identify barriers hampering efficient transduction and to develop an optimized AAV variant for EC transduction, we screened an AAV serotype 2-based peptide display library on primary human macrovascular EC. Using a new high-throughput selection and monitoring protocol, we identified a capsid variant, AAV-V EC, which outperformed the parental serotype as well as first-generation targeting vectors in EC transduction. AAV vector uptake was improved, resulting in significantly higher transgene expression levels from single-stranded vector genomes detectable within a few hours post-transduction. Notably, AAV-V EC transduced not only proliferating EC but also quiescent EC, although higher particle-per-cell ratios had to be applied. Also, induced pluripotent stem cell-derived endothelial progenitor cells, a novel tool in regenerative medicine and gene therapy, were highly susceptible toward AAV-V EC transduction. Thus, overcoming barriers by capsid engineering significantly expands the AAV tool kit for a wide range of applications targeting EC.

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KW - endothelial cells

KW - induced pluripotent stem cells

KW - uncoating

KW - uptake

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DO - 10.1089/hum.2019.027

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JF - Human Gene Therapy

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Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells

Abstract

Bibliographical note

Keywords

UN SDGs

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