epidermal-growth-factor and Peritoneal-Neoplasms

Potency, immunogenicity, and toxicity are three problems that limit the use of targeted toxins in solid tumour therapy.. To address potency, we used genetic engineering to develop a novel bispecific ligand-directed toxin (BLT) called EGF4KDEL, a novel recombinant anti-mesothelioma agent created by linking human epidermal growth factor (EGF) and interleukin-4 (IL-4) to truncated pseudomonas exotoxin (PE38) on the same single-chain molecule. Immunogenicity was reduced by mutating seven immunodominant B-cell epitopes on the PE38 molecule to create a new agent, EGF4KDEL 7Mut.. In vitro, bispecific EGF4KDEL showed superior anti-mesothelioma activity compared with its monospecific counterparts. Toxicity in mice was diminished by having both ligands on the same molecule, allowing administration of a 10-fold greater dose of BLT than a mixture of monomeric IL4KDEL and EGFKDEL. EGF4KDEL 7Mut, retained all of its functional activity and induced about 87% fewer anti-toxin antibodies than mice given the parental, non-mutated form. In vivo, intraperitoneal (IP) injection of the BLT showed significant (P<0.01) and impressive effects against two aggressive, malignant IP mesothelioma models when treatment was begun 14-16 days post tumour innoculation.. These data show that EGF4KDEL 7Mut is a promising new anti-mesothelioma agent that was developed to specifically address the obstacles facing clinical utility of targeted toxins.

Topics: ADP Ribose Transferases; Animals; Bacterial Toxins; Cell Line, Tumor; Epidermal Growth Factor; Exotoxins; Female; Humans; Immunotoxins; Interleukin-4; Male; Mesothelioma; Mice; Mice, Inbred BALB C; Peritoneal Neoplasms; Pseudomonas aeruginosa Exotoxin A; Recombinant Proteins; Virulence Factors; Xenograft Model Antitumor Assays

Although the association between asbestos exposure and mesothelioma development has been established for decades, very little is known regarding the molecular mechanism(s) by which asbestos fibers induce this disease. In this series of experiments, the potential for transforming growth factor alpha (TGF-alpha) to act as an autocrine growth factor in transformed mesothelial cells was examined in rats, a model system frequently used to assess the tumorigenic potential of fibrous particulates. Both asbestos-transformed cells and spontaneously transformed cells expressed functional EGF receptors, although only the asbestos-transformed cells expressed TGF-alpha. Expression of TGF-alpha transcripts was correlated with secretion of picogram amounts of growth factor into conditioned medium by the asbestos-transformed cells. In addition, whereas TGF-alpha inhibited the growth of spontaneously transformed mesothelial cells, it stimulated the growth of asbestos-transformed cells. Neutralizing antibody that recognized TGF-alpha secreted by the asbestos-transformed cells was able to inhibit the growth of these cells. Taken together, these data indicate that TGF-alpha acts as an autocrine growth factor for asbestos-transformed rat mesothelial cells. Therefore, in asbestos-transformed mesothelial cells, altered production and responsiveness to TGF-alpha distinguish these cells from spontaneously transformed mesothelial cells. These data suggest that differences in mesothelioma etiology may be reflected in differences in the molecular alterations present in these tumors.

Topics: Animals; Asbestos, Crocidolite; Blotting, Northern; Cell Division; Cell Line; Cell Line, Transformed; Epidermal Growth Factor; ErbB Receptors; Mesothelioma; Peritoneal Neoplasms; Phosphoproteins; Phosphotyrosine; Radioimmunoassay; Rats; Transforming Growth Factor alpha; Tumor Cells, Cultured; Tyrosine

Indirect evidence suggests that gonadal steroids and gonadotropins may have a role in the genesis of epithelial ovarian cancer. In the studies reported herein, we established 17 beta-estradiol (E2) secreting cell cultures from an omental metastasis of an epithelial ovarian cancer. We demonstrate that human chorionic gonadotropin (hCG), human follicle-stimulating hormone, and epidermal growth factor (EGF) increased cell growth in a dose- and time-dependent manner, whereas E2 inhibited cell growth in the nanomolar range. Epidermal growth factor was able to partially block the negative effect of E2; a similar but quantitatively lesser effect was observed with hCG. These results provide direct evidence to support the view that gonadotropins, EGF, TGF beta (transforming growth factor), and estradiol may modulate growth of metastatic epithelial ovarian cancer cells.

Topics: Cell Count; Cell Division; Chorionic Gonadotropin; Cyclic AMP; Cystadenocarcinoma; Dose-Response Relationship, Drug; Drug Interactions; Epidermal Growth Factor; Estradiol; Female; Follicle Stimulating Hormone; Humans; Middle Aged; Omentum; Ovarian Neoplasms; Peritoneal Neoplasms; Time Factors; Tumor Cells, Cultured

The present immunohistochemical studies used polyclonal antibodies specific to epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), and monoclonal antibodies to the extracellular binding domain of EGF receptor to elucidate their presence and cellular distribution in surgically induced endometriosis in the rat. Uterine segments were implanted in the mesenteric region adjacent to a blood vessel for a period of 4-6 weeks. During operative reexploration, the implanted tissues were intact, morphologically similar to the controls, and consisted of a cyst containing clear fluid and associated adhesion formation. All the uterine cell types immunostained with antibody to EGF. The highest immunostaining intensity was associated with inflammatory cells infiltrated among endometrial stromal cells, followed by luminal and glandular epithelial and stromal cells. The cysts consisted of an inner simple columnar epithelium, surrounded by several layers of smooth muscle and connective tissue. The cyst epithelial layer immunostained weakly, while the supporting wall stained moderately. The inflammatory cells found within the cyst cavity immunostained very intensely, comparable to those in the endometrium. The patterns of immunostaining for TGF-alpha and EGF receptor were similar to that observed for EGF. Myometrial smooth-muscle cells and endothelial and smooth-muscle cells of arterioles were also immunostained for EGF, TGF-alpha, and EGF receptor, but with lower intensity than that of the endometrium. These observations suggest that, like normal uterine tissue, endometrial implants produce EGF and TGF-alpha locally and contain receptors for EGF. These results imply a possible paracrine or autocrine role for growth factors in the establishment and/or maintenance of endometriotic tissue.

Topics: Animals; Cysts; Endometriosis; Endometrium; Epidermal Growth Factor; Epithelium; ErbB Receptors; Female; Immunohistochemistry; Mesentery; Muscle, Smooth; Myometrium; Peritoneal Neoplasms; Rats; Rats, Inbred Strains; Transforming Growth Factor alpha; Uterus