Adenovirus vector-based purging of multiple myeloma cells.
Adenoviruses are efficient gene delivery agents for a variety of neoplasms. In the present study, we have investigated the use of adenoviruses for the delivery of the thymidine kinase (tk) gene into multiple myeloma (MM) cells. We first demonstrated that MM cell lines and MM patient cells express both adenovirus receptors as well as the DF3/MUC1 protein, thus providing a rationale for using adenoviruses to selectively deliver genes under the control of the DF3 promoter. By using an adenoviral construct containing beta-galactosidase (beta-gal) gene driven by the DF3 promoter (Ad. DF3-betagal), we demonstrate greater than 80% transduction efficiency in OCI-My5 and RPMI 8226 MM cell lines at a multiplicity of infection of 1 to 100. Importantly, transduction with the tk gene driven by the DF3 promoter (Ad.DF3-tk) followed by treatment with 50 micromol/L ganciclovir (GCV) purged >/=6 log of contaminating OCI-My5 and RPMI 8226 MM cells within bone marrow mononuclear cells. In contrast, normal human hematopoietic progenitor cell number was unaffected under these conditions. Selectivity of DF3/MUC1 promoter was further confirmed, because Ad.DF3-betagal or Ad.DF3-tk did not transduce MUC1-negative HeLa cervical carcinoma cells. In addition, GCV treatment of Ad.DF3-tk-transduced RPMI 8226 MM cells did not induce a significant bystander effect. These findings demonstrate that transduction with Ad vectors using a tumor-selective promoter provides a highly efficient and selective approach for the ex vivo purging of MM cells.
Teoh
G
G
Department of Adult Oncology, Dana-Farber Cancer Institute, Boston, MA; and the Department of Haematology, Singapore General Hospital, Singapore.
Chen
L
L
Urashima
M
M
Tai
Y T
YT
Celi
L A
LA
Chen
D
D
Chauhan
D
D
Ogata
A
A
Finberg
R W
RW
Webb
I J
IJ
Kufe
D W
DW
Anderson
K C
KC
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D017502:Adenovirus E2 Proteins / Q000235:genetics; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004279:DNA, Viral / Q000032:analysis; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000821:virology; D006801:Humans; D016023:Integrins / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D018396:Mucin-1 / Q000096:biosynthesis / Q000235:genetics; D009101:Multiple Myeloma / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D019030:Receptors, Vitronectin; D013379:Substrate Specificity; D013937:Thymidine Kinase / Q000235:genetics; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D017502:Adenovirus E2 Proteins; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004279:DNA, Viral; D016023:Integrins; D018396:Mucin-1; D011991:Receptors, Virus; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; D013937:Thymidine Kinase; D001616:beta-Galactosidase; D015774:Ganciclovir