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.
| Language | English |
|---|---|
| Pages | 1284-1296 |
| Number of pages | 13 |
| Journal | Human Gene Therapy |
| Volume | 30 |
| Issue number | 10 |
| Early online date | 13 Aug 2019 |
| DOIs | |
| Publication status | Published - 23 Sep 2019 |
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Keywords
- AAV vectors
- endothelial cells
- induced pluripotent stem cells
- uncoating
- uptake
Cite this
}
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 journal › Article
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 -