Improved gene transfer efficiency to primary and established human pancreatic carcinoma target cells via epidermal growth factor receptor and integrin-targeted adenoviral vectors.
In this study we analyzed two ways of retargeting of Ad-vectors to human pancreatic carcinoma with the aim of enhancing the gene transfer efficiency. First, we analyzed the expression of the epidermal growth factor receptor (EGFR) on primary, as well as established pancreatic carcinoma cells by flow cytometry which revealed high expression levels of EGFR on the surface of these cells. We showed that EGFR-retargeted entry pathway using a bispecific fusion protein formed by a recombinant soluble form of truncated Coxsackie and Adenovirus Receptor (sCAR) genetically fused with human EGF (sCAR-EGF) redirects them to the EGFR leading to an enhanced gene transfer efficiency to pancreatic carcinoma cells. Since flow cytometry revealed absence of CAR expression, but the presence of at least one of both alphav integrins on the pancreatic carcinoma cells, a second way of targeting was investigated using a genetically modified Ad vector which has an RGD (Arg-Gly-Asp)-containing peptide inserted into the HI-loop of the fiber knob. This RGD targeted Ad (AdlucRGD) revealed efficient CAR-independent infection by allowing binding to cellular integrins resulting in a dramatic enhancement of gene transfer. These findings have direct relevance for Ad-vector based gene therapy strategies for pancreatic carcinoma.
Wesseling
J G
JG
Department of Experimental Hepatology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Bosma
P J
PJ
Krasnykh
V
V
Kashentseva
E A
EA
Blackwell
J L
JL
Reynolds
P N
PN
Li
H
H
Parameshwar
M
M
Vickers
S M
SM
Jaffee
E M
EM
Huibregtse
K
K
Curiel
D T
DT
Dmitriev
I
I
D000256:Adenoviridae / Q000235:genetics; D066246:ErbB Receptors / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D016023:Integrins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D019030:Receptors, Vitronectin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D016023:Integrins; D009363:Neoplasm Proteins; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; D066246:ErbB Receptors