Enhancement of gene transfer efficiency into human cancer cells by modification of retroviral vectors and addition of chemicals.

2000 ◽  
Author(s):  
J J Song ◽  
J H Kim ◽  
H Lee ◽  
E Kim ◽  
J Kim ◽  
...  
2008 ◽  
Vol 26 (1) ◽  
pp. 28-34 ◽  
Author(s):  
Yueping Li ◽  
Shudong Qiu ◽  
Liping Song ◽  
Qingfeng Yan ◽  
Guangxiao Yang

Oncogene ◽  
1999 ◽  
Vol 18 (13) ◽  
pp. 2189-2199 ◽  
Author(s):  
Takuya Fukazawa ◽  
Toshiyoshi Fujiwara ◽  
Yoshinori Morimoto ◽  
Jianghua Shao ◽  
Masahiko Nishizaki ◽  
...  

2010 ◽  
Vol 31 (11) ◽  
pp. 1822-1832 ◽  
Author(s):  
Christopher Gerner ◽  
Verena J. Haudek-Prinz ◽  
Andreas Lackner ◽  
Annemarie Losert ◽  
Barbara Peter-Vörösmarty ◽  
...  

Blood ◽  
1997 ◽  
Vol 90 (11) ◽  
pp. 4638-4645 ◽  
Author(s):  
Hans-Peter Kiem ◽  
Scott Heyward ◽  
Aaron Winkler ◽  
Jennifer Potter ◽  
James M. Allen ◽  
...  

Many diseases might be treated by gene therapy targeted to the hematopoietic system, but low rates of gene transfer achieved in humans and large animals have limited the application of this technique. We have developed a competitive hematopoietic repopulation assay in baboons to evaluate methods for improving gene transfer and have used this method to compare gene transfer rates for retroviral vectors having an envelope protein (pseudotype) from amphotropic murine retrovirus with similar vectors having an envelope protein derived from gibbon ape leukemia virus (GALV). We hypothesized that vectors with a GALV pseudotype might perform better based on our previous work with cultured human hematopoietic cells. CD34+ marrow cells from each of four untreated baboons were divided into two equal portions that were cocultivated for 48 hours with packaging cells producing equivalent titers of either amphotropic or GALV pseudotyped vectors containing the neo gene. The vectors contained small sequence differences to allow differentiation of cells genetically marked by the different vectors. Nonadherent and adherent cells from the cultures were infused into animals after they received a myeloablative dose of total body irradiation. Polymerase chain reaction (PCR) analysis for neo gene-specific sequences in colony-forming unit–granulocyte-macrophage from cell populations used for transplant showed gene transfer rates of 2.7%, 7.1%, <15%, and 3.9% with the amphotropic vectors and 7.1%, 11.3%, <15%, and 26.4% with the GALV pseudotyped vector. PCR analysis of peripheral blood and marrow cells after engraftment showed the neo gene to be present in all four animals analyzed at levels between 0.1% and 5%. Overall gene transfer efficiency was higher with the GALVpseudotyped vector than with the amphotropic vectors. Southern blot analysis in one animal confirmed a gene transfer efficiency of between 1% and 5%. The higher gene transfer efficiency with the GALV-pseudotyped vector correlated with higher levels of GALV receptor RNA compared with the amphotropic receptor in CD34+ hematopoietic cells. These results show that GALV-pseudotyped vectors are capable of transducing baboon marrow repopulating cells and may allow more efficient gene transfer rates for human gene therapy directed at hematopoietic cells. In addition, our data show considerable differences in gene transfer efficiency between individual baboons, suggesting that a competitive repopulation assay will be critical for evaluation of methods designed to improve gene transfer into hematopoietic stem cells.


2010 ◽  
Vol 4 (10-11) ◽  
pp. 867-867
Author(s):  
Christopher Gerner ◽  
Verena J. Haudek-Prinz ◽  
Andreas Lackner ◽  
Annemarie Losert ◽  
Barbara Peter-Vörösmarty ◽  
...  

Blood ◽  
1997 ◽  
Vol 90 (11) ◽  
pp. 4638-4645 ◽  
Author(s):  
Hans-Peter Kiem ◽  
Scott Heyward ◽  
Aaron Winkler ◽  
Jennifer Potter ◽  
James M. Allen ◽  
...  

Abstract Many diseases might be treated by gene therapy targeted to the hematopoietic system, but low rates of gene transfer achieved in humans and large animals have limited the application of this technique. We have developed a competitive hematopoietic repopulation assay in baboons to evaluate methods for improving gene transfer and have used this method to compare gene transfer rates for retroviral vectors having an envelope protein (pseudotype) from amphotropic murine retrovirus with similar vectors having an envelope protein derived from gibbon ape leukemia virus (GALV). We hypothesized that vectors with a GALV pseudotype might perform better based on our previous work with cultured human hematopoietic cells. CD34+ marrow cells from each of four untreated baboons were divided into two equal portions that were cocultivated for 48 hours with packaging cells producing equivalent titers of either amphotropic or GALV pseudotyped vectors containing the neo gene. The vectors contained small sequence differences to allow differentiation of cells genetically marked by the different vectors. Nonadherent and adherent cells from the cultures were infused into animals after they received a myeloablative dose of total body irradiation. Polymerase chain reaction (PCR) analysis for neo gene-specific sequences in colony-forming unit–granulocyte-macrophage from cell populations used for transplant showed gene transfer rates of 2.7%, 7.1%, <15%, and 3.9% with the amphotropic vectors and 7.1%, 11.3%, <15%, and 26.4% with the GALV pseudotyped vector. PCR analysis of peripheral blood and marrow cells after engraftment showed the neo gene to be present in all four animals analyzed at levels between 0.1% and 5%. Overall gene transfer efficiency was higher with the GALVpseudotyped vector than with the amphotropic vectors. Southern blot analysis in one animal confirmed a gene transfer efficiency of between 1% and 5%. The higher gene transfer efficiency with the GALV-pseudotyped vector correlated with higher levels of GALV receptor RNA compared with the amphotropic receptor in CD34+ hematopoietic cells. These results show that GALV-pseudotyped vectors are capable of transducing baboon marrow repopulating cells and may allow more efficient gene transfer rates for human gene therapy directed at hematopoietic cells. In addition, our data show considerable differences in gene transfer efficiency between individual baboons, suggesting that a competitive repopulation assay will be critical for evaluation of methods designed to improve gene transfer into hematopoietic stem cells.


2003 ◽  
Vol 103 (6) ◽  
pp. 833-839 ◽  
Author(s):  
Fuminori Teraishi ◽  
Yoshihiko Kadowaki ◽  
Yasuhisa Tango ◽  
Takeshi Kawashima ◽  
Tatsuo Umeoka ◽  
...  

Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
S Nam ◽  
R Buettner ◽  
X Liu ◽  
J Turkson ◽  
D Kim ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document