ABSTRACT
BACKGROUND: Recombinant vesicular stomatitis virus expressing interferon-ß (VSV-IFN-ß) has demonstrated antitumor activity in vitro and in vivo. In preparation for clinical testing in human squamous cell carcinoma (SCC) of the head and neck, we conducted preclinical studies of VSV-IFN-ß in syngeneic SCC models. METHODS: In vitro, VSV-IFN-ß (expressing rat or mouse interferon [IFN]-ß)-induced cytotoxicity and propagated in rat (FAT-7) or mouse (SCC-VII) SCC cells during normoxia and hypoxia. In vivo, intratumoral administration of VSV-rat-IFN-ß or VSV-human-IFN-ß in FAT-7 bearing or non-tumor bearing immunocompetent rats did not result in acute organ toxicity or death. RESULTS: VSV-r-IFN-ß replicated predominantly in tumors and a dose dependent anti-VSV antibody response was observed. Intratumoral or intravenous administration of VSV-IFN-ß resulted in growth delay and improved survival compared with controls. CONCLUSION: The above data confirm safety and feasibility of VSV-IFN-ß administration in immunocompetent animals and support its clinical evaluation in advanced human head and neck cancer.
Subject(s)
Carcinoma, Squamous Cell/therapy , Head and Neck Neoplasms/therapy , Interferon-beta/pharmacology , Oncolytic Virotherapy/methods , Vesiculovirus/isolation & purification , Analysis of Variance , Animals , Blotting, Western , Carcinoma, Squamous Cell/immunology , Carcinoma, Squamous Cell/pathology , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Feasibility Studies , Female , Head and Neck Neoplasms/immunology , Head and Neck Neoplasms/pathology , Humans , Immunocompetence , Kaplan-Meier Estimate , Mice , Random Allocation , Rats , Rats, Inbred F344 , Safety , Statistics, Nonparametric , Transplantation, Isogeneic , Treatment Outcome , Tumor Cells, Cultured , Vesicular Stomatitis/virologyABSTRACT
PURPOSE: Detect tumor-related DNA using LigAmp in histologically clear margins and associate results with clinical outcome. EXPERIMENTAL DESIGN: Patients with head and neck cancer were registered for molecular analysis of surgical margins. Adequacy of resection was ensured using histologic margin analysis. Further margins were then harvested and DNA extracted. TP53 mutations in tumor were determined using Affymetrix p53 GeneChip. Margins were analyzed by Ligamp in comparison with standard curves for quantification of mutant DNA. Ligation used two oligonucleotides to isolate DNA targeting the mutation. Ligated DNA was amplified using real-time PCR. The quantity of mutation in the margin was determined as percent of mutant species relative to plasmid and relative to tumor. Cutpoints were identified and defined groups were evaluated for local failure-free, cancer-specific, and overall survival. Study margins were examined for presence of tumor by light microscopy. RESULTS: Tissue from 95 patients with common mutations was analyzed. Fifteen experienced local recurrence. Cutpoints of 0.15% for mutant species relative to plasmid and 0.5% for mutant species relative to tumor were chosen as most selective of recurrent cases. LigAmp had slightly better area under the receiver operator characteristic curve (P = 0.09) than light microscopy correctly predicting 9 of 15 recurrent tumors. There were 6 false negative cases and 26 false positive results. No statistically significant distinctions were observed in cancer-specific or overall survival in this limited cohort. CONCLUSIONS: Ligamp provides quantifiable, sensitive detection of mutant DNA in histologically normal margins. Detection of mutant species in margins may identify patients at risk of local recurrence. (Clin Cancer Res 2009;15(24):7658-65).
ABSTRACT
BACKGROUND: The abrogation of function of the tumor-suppressor protein p53 as a result of mutation of its gene, TP53, is one of the most common genetic alterations in cancer cells. We evaluated TP53 mutations and survival in patients with squamous-cell carcinoma of the head and neck. METHODS: A total of 560 patients with squamous-cell carcinoma of the head and neck who were treated surgically with curative intent were enrolled in our prospective multicenter, 7-year study. TP53 mutations were analyzed in DNA from the tumor specimens with the use of the Affymetrix p53 chip and the Surveyor DNA endonuclease and denaturing high-performance liquid chromatography. Mutations were classified into two groups, disruptive and nondisruptive, according to the degree of disturbance of protein structure predicted from the crystal structure of the p53-DNA complexes. TP53 mutational status was compared with clinical outcome. RESULTS: TP53 mutations were found in tumors from 224 of 420 patients (53.3%). As compared with wild-type TP53, the presence of any TP53 mutation was associated with decreased overall survival (hazard ratio for death, 1.4; 95% confidence interval [CI], 1.1 to 1.8; P=0.009), with an even stronger association with disruptive mutations (hazard ratio, 1.7; 95% CI, 1.3 to 2.4; P<0.001) and no significant association with nondisruptive mutations (hazard ratio, 1.2; 95% CI, 0.9 to 1.7; P=0.16). In multivariate analyses a disruptive TP53 alteration, as compared with the absence of a TP53 mutation, had an independent, significant association with decreased survival (hazard ratio, 1.7; 95% CI, 1.2 to 2.4; P=0.003). CONCLUSIONS: Disruptive TP53 mutations in tumor DNA are associated with reduced survival after surgical treatment of squamous-cell carcinoma of the head and neck.
