Mesothelin-mediated targeting of adenoviral vectors for ovarian cancer gene therapy.
Adenoviruses (Ads) are efficient gene transfer vehicles, but Ad-mediated gene therapy for ovarian cancer remains limited in vivo by inefficient and nonspecific gene transfer. Mesothelin (MSLN), a cell surface glycoprotein, is overexpressed in ovarian cancer but not in normal tissues except mesothelial cells. Therefore, MSLN is an attractive candidate for transcriptional and transductional targeting in the context of ovarian cancer gene therapy. We evaluated the expression of MSLN mRNA and MSLN surface protein in ovarian cancer cells. Ads containing the MSLN promoter driving reporter gene expression were created and tested in ovarian cancer cell lines and purified ovarian cancer cells isolated from patients. To evaluate transductional targeting, we used an Ad vector containing an Fc-binding domain within the fiber protein, which served as a docking domain for binding with anti-MSLN immunoglobulins. Both RT-PCR and flow cytometry revealed high MSLN gene and protein expression in ovarian cancer cells. The MSLN promoter was activated in ovarian cancer cells, but showed significantly reduced activity in normal control cells. Transductional targeting of Ads via anti-MSLN antibody increased transgene expression in ovarian cancer cells. This report describes the use of MSLN for transcriptional as well as transductional targeting strategies for ovarian cancer gene therapy.
Breidenbach
M
M
Division of Human Gene Therapy, Departments of Medicine, Surgery, Pathology and the Gene Therapy Center, Birmingham, AL 35294-2172, USA.
Rein
D T
DT
Everts
M
M
Glasgow
J N
JN
Wang
M
M
Passineau
M J
MJ
Alvarez
R D
RD
Korokhov
N
N
Curiel
D T
DT
D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D058851:GPI-Linked Proteins; D015870:Gene Expression; D018390:Gene Targeting; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics*; D000090204:Mesothelin; D010051:Ovarian Neoplasms / Q000628:therapy*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D014161:Transduction, Genetic / Q000379:methods; D014407:Tumor Cells, Cultured
D058851:GPI-Linked Proteins; C000715527:MSLN protein, human; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D000090204:Mesothelin