Tumor endothelial marker 7 (TEM-7): a novel target for antiangiogenic therapy.

Antiangiogenesis has been validated as a therapeutic strategy to treat cancer, however, a need remains to identify new targets and therapies for specific diseases and to improve clinical benefit from antiangiogenic agents. Tumor endothelial marker 7 (TEM-7) was investigated as a possible target for therapeutic antiangiogenic intervention in cancer. TEM-7 expression was assessed by in situ hybridization or by immunohistochemistry (IHC) in 130 formalin-fixed paraffin-embedded (FFPE) and 410 frozen human clinical specimens of cancer plus 301 normal tissue samples. In vitro TEM-7 expression was evaluated in 4 human endothelial cell models and in 32 human cancer cell lines by RT-PCR and flow cytometry. An anti-TEM-7 antibody was tested in vitro on human SKOV3 ovarian and MDA-MB-231 breast carcinoma cells that expressed TEM-7 in antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis assays. In frozen tumor tissues, TEM-7 mRNA and protein was detected in all but one of the cancer types tested and was infrequently expressed in normal frozen tissues. In FFPE tumor tissues, TEM-7 protein was detected by IHC in colon, breast, lung, bladder, ovarian and endometrial cancers and in sarcomas. TEM-7 protein was not detected in head and neck, prostate or liver cancers. TEM-7 expression was restricted to the vasculature and was absent from tumor cells. In vitro, TEM-7 was not detected in human microvascular endothelial cells (HMVEC) or human umbilical vein endothelial cells (HUVEC) but was induced in endothelial precursor/progenitor cells (EPC) in the presence of the mitogen phorbol ester PMA. An anti-TEM-7 antibody mediated ADCC and phagocytosis in SKOV3 and MDA-MB-231 cell lines infected with an adenovirus expressing TEM-7. These data demonstrate that TEM-7 is a vascular protein associated with angiogenic states. TEM-7 is a novel and attractive target for antiangiogenic therapy.

Endosialin protein expression and therapeutic target potential in human solid tumors: sarcoma versus carcinoma.

PURPOSE: Endosialin/CD248/tumor endothelial marker 1 is expressed in stromal cells, endothelial cells, and pericytes in various tumors; however, few studies have focused on expression in malignant cells. EXPERIMENTAL DESIGN: We studied expression of endosialin in clinical specimens, cell culture, and animal models and designed an anti-endosialin therapeutic prototype. RESULTS: Fifty human tumor cell lines and 6 normal cell types in culture were assayed by reverse transcription-PCR and/or flow cytometry for endosialin. Cell surface protein was found on 7 sarcoma lines, 1 neuroblastoma, and 4 normal cell types in culture. A fully human anti-endosialin antibody bound to human A-673 Ewing's sarcoma cells and SK-N-AS neuroblastoma cells but not HT-1080 cells. Exposure of cells to an anti-human IgG conjugated to saporin resulted in growth inhibition only of endosialin-expressing cells. Endosialin expression was assessed by immunohistochemistry in 250 clinical specimens of human cancer including 20 cancer subtypes. Endosialin is frequently found in human cancers. Endosialin expression is mainly a perivascular feature in carcinomas, with some expression in stromal cells. In sarcomas, endosialin is expressed by malignant cells, perivascular cells, and stromal cells. Development and characterization of experimental models for studying endosialin biology in sarcomas and evaluating anti-endosialin therapies is presented. CONCLUSIONS: Findings suggest that an anti-endosialin immunotoxin might be a promising therapeutic approach for endosialin-positive neoplasia, especially synovial sarcoma, fibrosarcoma, malignant fibrous histiocytoma, liposarcoma, and osteosarcoma. Thus, a diagnostic/therapeutic targeted therapeutic approach to treatment of endosialin-expressing tumors may be possible.

Protective effect of glycine in mesenteric ischemia and reperfusion injury in a rat model.

PURPOSE: Glycine has a protective effect in renal and skeletal muscle ischemia. The purpose of this study was to evaluate the effect of glycine in mesenteric ischemia and reperfusion injury in a rat model. METHODS: Twenty-four anesthetized male Sprague-Dawley rats were subjected to 1 hour of mesenteric ischemia followed by 2 hours of reperfusion. Control animals received normal saline solution intravenously at 0.01 mL/g of body weight/h during ischemia and reperfusion. Treated animals received glycine at 0.5, 0.75, or 1.0 mg/g of body weight, dissolved in saline solution and infused at 0.01 mL/g/h for 2 hours. Animals were killed at the end of the experiment, and proximal, middle, and distal segments of the small bowel were isolated. Sections of the segments stained with hematoxylin-eosin were subjected to histologic examination (as per modified Chiu grading system) and morphometric analysis consisting of measurement of bowel wall, muscularis and mucosal thickness, epithelial coverage, and villar circumference. Isometric tension responses to electrical stimulation (10, 30, 50, 100 Hz), high doses of potassium (120 mmol/L), and carbachol (0.1, 0.5, 1.0, 5.0 micromol/L) were recorded in a multimuscle chamber. Statistical analysis was performed with unpaired t test and one-way analysis of variance. RESULTS: The middle and distal segments of the small bowel in glycine-treated animals showed better histologic grade compared with saline solution-treated control rats (P <.05). At morphometric analysis, total thickness, mucosal thickness, and villar circumference ratio were well preserved in the middle and distal segments of the small bowel in the glycine-treated group (P <.05). No significant differences were observed in the proximal bowel segments between glycine-treated and control animals, because the proximal segment was not subjected to much ischemia. No differences were noted in percentage of epithelial coverage. Isometric tension responses evoked by electrical stimulation were greater (P <.05) in the middle and distal segments treated with glycine as compared with control segments. Carbachol-evoked contractions were stronger (P <.05) in the small bowel segments of animals treated with glycine. The responses evoked by 120 mmol/L of potassium were stronger in the distal segments of the small bowel in the glycine-treated group (P <.05). This cytoprotective effect of glycine was not dose-dependent. CONCLUSIONS: Glycine improved mucosal viability in the ischemia and reperfusion injury rat model. Mucosal thickness and villous circumference ratio were reliable objective parameters for evaluation of intestinal ischemia injury. Glycine improved the contractile responses of the bowel segments also, probably by altering the physiologic mechanisms underlying force generation. Further studies are required to elucidate the mechanism of the cytoprotective action of glycine.