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

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

LanguageEnglish
Pages1284-1296
Number of pages13
JournalHuman Gene Therapy
Volume30
Issue number10
Early online date13 Aug 2019
DOIs
Publication statusPublished - 23 Sep 2019

Fingerprint

Dependovirus
Capsid
Endothelial Cells
Regenerative Medicine
Genetic Therapy
Peptide Library
Induced Pluripotent Stem Cells
Transgenes
Genome

Keywords

  • AAV vectors
  • endothelial cells
  • induced pluripotent stem cells
  • uncoating
  • uptake

Cite this

Zhang, L., Rossi, A., Lange, L., Meumann, N., Koitzsch, U., Christie, K., ... 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
Zhang, L ; Rossi, A ; Lange, L ; Meumann, N ; Koitzsch, U ; Christie, Kathleen ; Nesbit, M. Andrew ; Moore, Tara C. B. ; Hacker, U.T. ; Morgan, M. ; Hoffmann, D. ; Zengel, James ; Carette, Jan E. ; Schambach, A. ; Salvetti, A. ; Odenthal, Margarete ; Büning, Hildegard. / Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells. In: Human Gene Therapy. 2019 ; Vol. 30, No. 10. pp. 1284-1296.
@article{2addd54f257942edb7095dd414254c80,
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",
year = "2019",
month = "9",
day = "23",
doi = "10.1089/hum.2019.027",
language = "English",
volume = "30",
pages = "1284--1296",
journal = "Human Gene Therapy",
issn = "1043-0342",
number = "10",

}

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

Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells. / Zhang, L; Rossi, A; Lange, L; Meumann, N; Koitzsch, U; Christie, Kathleen; Nesbit, M. Andrew; Moore, Tara C. B.; Hacker, U.T.; Morgan, M.; Hoffmann, D.; Zengel, James; Carette, Jan E.; Schambach, A.; Salvetti, A.; Odenthal, Margarete; Büning, Hildegard.

In: Human Gene Therapy, Vol. 30, No. 10, 23.09.2019, p. 1284-1296.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells

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

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.

KW - AAV vectors

KW - endothelial cells

KW - induced pluripotent stem cells

KW - uncoating

KW - uptake

UR - http://www.scopus.com/inward/record.url?scp=85072848537&partnerID=8YFLogxK

U2 - 10.1089/hum.2019.027

DO - 10.1089/hum.2019.027

M3 - Article

VL - 30

SP - 1284

EP - 1296

JO - Human Gene Therapy

T2 - Human Gene Therapy

JF - Human Gene Therapy

SN - 1043-0342

IS - 10

ER -