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Deficiency of oncoretrovirally transduced hematopoietic stem cells and correction through ex vivo expansion.

Bryder, David LU ; Sasaki, Yutaka LU and Jacobsen, Sten Eirik W LU (2005) In Journal of Gene Medicine 7(2). p.137-144
Abstract
Background Extensive efforts to develop hematopoietic stem cell (HSC) based gene therapy have been hampered by low gene marking. Major emphasis has so far been directed at improving gene transfer efficiency, but low gene marking in transplanted recipients might equally well reflect compromised repopulating activity of transduced cells, competing for reconstitution with endogenous and unmanipulated stem cells. Methods The autologous settings of clinical gene therapy protocols preclude evaluation of changes in repopulating ability following transduction; however, using a congenic mouse model, allowing for direct evaluation of gene marking of lympho-myeloid progeny, we show here that these issues can be accurately addressed. Results We... (More)
Background Extensive efforts to develop hematopoietic stem cell (HSC) based gene therapy have been hampered by low gene marking. Major emphasis has so far been directed at improving gene transfer efficiency, but low gene marking in transplanted recipients might equally well reflect compromised repopulating activity of transduced cells, competing for reconstitution with endogenous and unmanipulated stem cells. Methods The autologous settings of clinical gene therapy protocols preclude evaluation of changes in repopulating ability following transduction; however, using a congenic mouse model, allowing for direct evaluation of gene marking of lympho-myeloid progeny, we show here that these issues can be accurately addressed. Results We demonstrate that conditions supporting in vitro stem cell self-renewal efficiently promote oncoretroviral-mediated gene transfer to multipotent adult bone marrow stem cells, without prior in vivo conditioning. Despite using optimized culture conditions, transduction resulted in striking losses of repopulating activity, translating into low numbers of gene marked cells in competitively repopulated mice. Subjecting transduced HSCs to an ex vivo expansion protocol following the transduction procedure could partially reverse this loss. Conclusions These studies suggest that loss of repopulating ability of transduced HSCs rather than low gene transfer efficiency might be the main problem in clinical gene therapy protocols, and that a clinically feasible ex vivo expansion approach post-transduction can markedly improve reconstitution with gene marked stem cells. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
transplantation, hematopoiesis, gene therapy
in
Journal of Gene Medicine
volume
7
issue
2
pages
137 - 144
publisher
John Wiley & Sons
external identifiers
  • pmid:15538726
  • wos:000227253900001
  • scopus:14844293836
ISSN
1521-2254
DOI
10.1002/jgm.658
language
English
LU publication?
yes
id
fda2d766-16d3-472e-b4b9-191878ed298d (old id 130881)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15538726&dopt=Abstract
date added to LUP
2007-07-23 16:01:12
date last changed
2017-01-01 04:37:02
@article{fda2d766-16d3-472e-b4b9-191878ed298d,
  abstract     = {Background Extensive efforts to develop hematopoietic stem cell (HSC) based gene therapy have been hampered by low gene marking. Major emphasis has so far been directed at improving gene transfer efficiency, but low gene marking in transplanted recipients might equally well reflect compromised repopulating activity of transduced cells, competing for reconstitution with endogenous and unmanipulated stem cells. Methods The autologous settings of clinical gene therapy protocols preclude evaluation of changes in repopulating ability following transduction; however, using a congenic mouse model, allowing for direct evaluation of gene marking of lympho-myeloid progeny, we show here that these issues can be accurately addressed. Results We demonstrate that conditions supporting in vitro stem cell self-renewal efficiently promote oncoretroviral-mediated gene transfer to multipotent adult bone marrow stem cells, without prior in vivo conditioning. Despite using optimized culture conditions, transduction resulted in striking losses of repopulating activity, translating into low numbers of gene marked cells in competitively repopulated mice. Subjecting transduced HSCs to an ex vivo expansion protocol following the transduction procedure could partially reverse this loss. Conclusions These studies suggest that loss of repopulating ability of transduced HSCs rather than low gene transfer efficiency might be the main problem in clinical gene therapy protocols, and that a clinically feasible ex vivo expansion approach post-transduction can markedly improve reconstitution with gene marked stem cells.},
  author       = {Bryder, David and Sasaki, Yutaka and Jacobsen, Sten Eirik W},
  issn         = {1521-2254},
  keyword      = {transplantation,hematopoiesis,gene therapy},
  language     = {eng},
  number       = {2},
  pages        = {137--144},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Gene Medicine},
  title        = {Deficiency of oncoretrovirally transduced hematopoietic stem cells and correction through ex vivo expansion.},
  url          = {http://dx.doi.org/10.1002/jgm.658},
  volume       = {7},
  year         = {2005},
}