Development of an ErbB-overexpressing A-431 optical reporting tumor xenograft model to assess targeted photodynamic therapy regimens.
To better assess the efficacy of erbB-targeted therapies, it would help to have optical reporting human tumor xenograft models that abundantly express erbB receptors. A-431 cells have frequently been used in erbB1-targeting studies, but a well-characterized optical reporting version of the cell line has not been readily available. In this study, optical reporting A-431 clones were developed that express both a fluorescent protein reporter (green, GFP; or red, RFP) and a bioluminescent reporter, firefly luciferase. Reporter genes were transduced into cells using commercial lentiviral vectors, and clonal selection was carried out using a series of procedures. A number of clones were isolated for further characterization. A GFP/luciferase clone, A-431/D4, and an RFP/luciferase clone, A-431/G4, were obtained that exhibit erbB1 expression levels and tumor growth kinetics similar to the parental cells. To demonstrate the utility of the optical reporting clones, A-431/G4 tumors were grown subcutaneously in nude mice and treated with vascular-targeted photodynamic therapy (PDT), which targets the angiogenic consequences of erbB signaling. The A-431/G4 tumor model permitted highly sensitive longitudinal monitoring of PDT treatment response using optical imaging. A-431/D4 and A-431/G4 optical reporting tumor models should also prove useful for assessing therapies that directly target the erbB1 receptor.
Savellano
Mark D
MD
Surgical Research Laboratories, Dartmouth Medical School, Lebanon, NH, USA. mark.savellano@dartmouth.edu
Owusu-Brackett
Nicci
N
Son
Ji
J
Callier
Thierri
T
Savellano
Dagmar Högemann
DH
D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D017930:Genes, Reporter; D018773:Genes, erbB-1; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000188:drug therapy* / Q000235:genetics*; D010778:Photochemotherapy / Q000295:instrumentation; D023041:Xenograft Model Antitumor Assays; D000097573:Red Fluorescent Protein
D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein