Antibody-directed double suicide gene therapy targeting of MUC1- positive leukemia cells in vitro and in vivo.
Our aim was to specifically transfer the cytosine deaminase (CD) and thymidine kinase (TK) genes into mucin 1 (MUC1)-positive leukemia cells by anti-MUC1 antibody directed infection of replication-defective lentivirus and to evaluate the targeted cytotoxicity of double suicide genes to leukemia. The target gene vector (containing CD and TK) and envelope (containing GFP and anti-MUC1) and packaging plasmids were cotransfected into 293T cells to produce the recombinant lentivirus. Suicide genes in virus-infected leukemia cells (U937, Jurkat, and K562) were detected by western blot. The cytotoxicity and bystander effect in vitro and the therapeutic effect in vivo were detected after treatment with the prodrugs. The results revealed that combined treatment with prodrug 5-fluorocytosine (5-FC) and ganciclovir (GCV) inhibited leukemia cell growth and caused significant bystander effect than treatment with either prodrug alone. TK/GCV treatment alone induced degeneration and cell death while the effect of CD/5-FC alone mainly caused vacuolar degeneration and necrosis. The addictive effects of combinatorial use of GCV and 5-FC mainly induced swelling of the mitochondria followed by necrosis of the leukemia cells. In vivo experiments revealed that both single and combinatorial prodrug treatments could prolong the survival time of leukemic mice. In summary, anti-MUC1 antibody directed lentiviral vector successfully transduced dual suicide genes and exerted targeted cytotoxicity against MUC1 positive leukemia cells. This targeted lentiviral dual suicide gene delivering system provides a promising approach for clinical treatment of leukemia in future.
Xiao-Ya
XY
Dong
Wen-Qian
WQ
Wang
Yu
Y
Zhao
Xu-Dong
XD
Li
Zhi-Gang
ZG
Fang
Dong-Jun
DJ
Lin
Ruo-Zhi
RZ
Xiao
Ren-Wei
RW
Huang
Guang-Jin
GJ
Pan
Hematological Department & Research Institute, Third Hospital of Sun Yat-sen University, Guangzhou, Guangdong, Tian-He Road 600, China 510630. jiajun.l@163.com.
Jia-Jun
JJ
Liu
D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation / Q000187:drug effects; D043525:Cytosine Deaminase / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005437:Flucytosine / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D018390:Gene Targeting / Q000379:methods; D018014:Gene Transfer Techniques; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D016086:Lentivirus / Q000235:genetics; D007938:Leukemia / Q000235:genetics* / Q000628:therapy*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D018396:Mucin-1 / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011355:Prodrugs / Q000494:pharmacology; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism; D020298:U937 Cells
C060192:MUC1 protein, human; D018396:Mucin-1; D011355:Prodrugs; D005437:Flucytosine; D013937:Thymidine Kinase; D043525:Cytosine Deaminase; D015774:Ganciclovir