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A retroviral packaging cell line for pseudotype vectors based on glioma-infiltrating progenitor cells.

Fischer, Yvonne LU ; Miletic, Hrvoje; Giroglou, Tsanan; Litwak, Sara; Stenzel, Werner; Neumann, Harald and von Laer, Dorothee (2007) In Journal of Gene Medicine 9(5). p.335-344
Abstract
Early clinical trials for gene therapy of human gliomas with retroviral packaging cells (PC) have been hampered by low transduction efficacy and lack of dissemination of PC within the tumor. In the current approach, these issues have been addressed by creating a stable packaging cell line for retroviral vectors pseudotyped with glycoproteins of lymphocytic choriomeningitis virus (LCMV) based on tumor-infiltrating progenitor cells. Methods Tumor-infiltrating progenitor cells, which had been isolated from adult rat bone marrow (BM-TIC), were modified to stably express Gag-Pol proteins of moloney murine leukemia virus (Mo-MLV) and glycoproteins of LCMV. Packaging of a retroviral vector was measured by titration experiments on human fibroblast... (More)
Early clinical trials for gene therapy of human gliomas with retroviral packaging cells (PC) have been hampered by low transduction efficacy and lack of dissemination of PC within the tumor. In the current approach, these issues have been addressed by creating a stable packaging cell line for retroviral vectors pseudotyped with glycoproteins of lymphocytic choriomeningitis virus (LCMV) based on tumor-infiltrating progenitor cells. Methods Tumor-infiltrating progenitor cells, which had been isolated from adult rat bone marrow (BM-TIC), were modified to stably express Gag-Pol proteins of moloney murine leukemia virus (Mo-MLV) and glycoproteins of LCMV. Packaging of a retroviral vector was measured by titration experiments on human fibroblast cells as well as on mouse and human glioma cell lines. Additionally, gene transfer was tested in a rat glioma model in vivo. Results The BM-TIC-derived packaging cell line (BM-TIPC) produced retroviral vectors with titers between 2-8 x 10(3) transducing units (TU)/ml. Extended culturing of BM-TIPC over several weeks and freezing/thawing of cells did not affect vector titers. No replication-competent retrovirus was released from BM-TIPC. In a rat glioma model, BM-TIPC infiltrated the tumors extensively and with high specificity. Moreover, BM-TIPC mediated transduction of glioma cells in vivo. Conclusion This proof-of-principle study shows that primary adult progenitor cells with tumor-infiltrating capacity can be genetically modified to stably produce retroviral LCMV pseudotype vectors. These BM-TIPC may be a useful tool to enhance specificity and efficacy of gene transfer to gliomas in patients. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Gene Medicine
volume
9
issue
5
pages
335 - 344
publisher
John Wiley & Sons
external identifiers
  • wos:000247051100002
  • scopus:34249034432
ISSN
1521-2254
DOI
10.1002/jgm.1032
language
English
LU publication?
yes
id
aadc09fc-0ec8-4e5a-8d29-e20e6cf54115 (old id 168480)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17474071&dopt=Abstract
date added to LUP
2007-07-23 16:19:58
date last changed
2017-01-01 04:26:33
@article{aadc09fc-0ec8-4e5a-8d29-e20e6cf54115,
  abstract     = {Early clinical trials for gene therapy of human gliomas with retroviral packaging cells (PC) have been hampered by low transduction efficacy and lack of dissemination of PC within the tumor. In the current approach, these issues have been addressed by creating a stable packaging cell line for retroviral vectors pseudotyped with glycoproteins of lymphocytic choriomeningitis virus (LCMV) based on tumor-infiltrating progenitor cells. Methods Tumor-infiltrating progenitor cells, which had been isolated from adult rat bone marrow (BM-TIC), were modified to stably express Gag-Pol proteins of moloney murine leukemia virus (Mo-MLV) and glycoproteins of LCMV. Packaging of a retroviral vector was measured by titration experiments on human fibroblast cells as well as on mouse and human glioma cell lines. Additionally, gene transfer was tested in a rat glioma model in vivo. Results The BM-TIC-derived packaging cell line (BM-TIPC) produced retroviral vectors with titers between 2-8 x 10(3) transducing units (TU)/ml. Extended culturing of BM-TIPC over several weeks and freezing/thawing of cells did not affect vector titers. No replication-competent retrovirus was released from BM-TIPC. In a rat glioma model, BM-TIPC infiltrated the tumors extensively and with high specificity. Moreover, BM-TIPC mediated transduction of glioma cells in vivo. Conclusion This proof-of-principle study shows that primary adult progenitor cells with tumor-infiltrating capacity can be genetically modified to stably produce retroviral LCMV pseudotype vectors. These BM-TIPC may be a useful tool to enhance specificity and efficacy of gene transfer to gliomas in patients.},
  author       = {Fischer, Yvonne and Miletic, Hrvoje and Giroglou, Tsanan and Litwak, Sara and Stenzel, Werner and Neumann, Harald and von Laer, Dorothee},
  issn         = {1521-2254},
  language     = {eng},
  number       = {5},
  pages        = {335--344},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Gene Medicine},
  title        = {A retroviral packaging cell line for pseudotype vectors based on glioma-infiltrating progenitor cells.},
  url          = {http://dx.doi.org/10.1002/jgm.1032},
  volume       = {9},
  year         = {2007},
}