Targeting tumor cells via EGF receptors: selective toxicity of an HBEGF-toxin fusion protein.
Over-expression of the epidermal growth factor receptor (EGFR) is a hallmark of numerous solid tumors, thus providing a means of selectively targeting therapeutic agents. Heparin-binding epidermal growth factor (HBEGF) binds to EGFRs with high affinity and to heparan sulfate proteoglycans, resulting in increased mitogenic potential compared to other EGF family members. We have investigated the feasibility of using HBEGF to selectively deliver a cytotoxic protein into EGFR-expressing tumor cells. Recombinant fusion proteins consisting of mature human HBEGF fused to the plant ribosome-inactivating protein saporin (SAP) were expressed in Escherichia coli. Purified HBEGF-SAP chimeras inhibited protein synthesis in a cell-free assay and competed with EGF for binding to receptors on intact cells. A construct with a 22-amino-acid flexible linker (L22) between the HBEGF and SAP moieties exhibited an affinity for the EGFR that was comparable to that of HBEGF. The sensitivity to HBEGF-L22-SAP was determined for a variety of human tumor cell lines, including the 60 cell lines comprising the National Cancer Institute Anticancer Drug Screen. HBEGF-L22-SAP was cytotoxic in vitro to a variety of EGFR-bearing cell lines and inhibited growth of EGFR-over-expressing human breast carcinoma cells in vivo. In contrast, the fusion protein had no effect on small-cell lung carcinoma cells, which are EGFR-deficient. Our results demonstrate that fusion proteins composed of HBEGF and SAP exhibit targeting specificity and cytotoxicity that may be of therapeutic value in treating a variety of EGFR-bearing malignancies.
Selective Genetics, Inc., San Diego, CA 92121, USA. loisc@selectivegenetics.com
L A
LA
Chandler
B A
BA
Sosnowski
J R
JR
McDonald
J E
JE
Price
S L
SL
Aukerman
A
A
Baird
G F
GF
Pierce
L L
LL
Houston
D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors / Q000378:metabolism*; D005240:Feasibility Studies; D005822:Genetic Vectors; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D000922:Immunotoxins / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism* / Q000494:pharmacology; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D014407:Tumor Cells, Cultured / Q000378:metabolism
C587883:HBEGF protein, human; D066257:Heparin-binding EGF-like Growth Factor; D000922:Immunotoxins; D036341:Intercellular Signaling Peptides and Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D009699:N-Glycosyl Hydrolases; D000076985:Saporins