Abstract 55: Programmable bio-nano-chip (p-BNC) based cytomorphometric and biomarker expression analysis and phenotypic characterization of Fanconi anemia oral cancer cells.
Authors: Vigneswaran Nadarajah , Floriano Pierre , Wu Jean , Wu Annie , Gillenwater Ann , McDevitt John 
Published: August 2014

Background: Fanconi anemia (FA) is an autosomal recessive disorder caused by mutations of genes in the FA pathway that are involved in the repair of DNA interstrand cross-links and DNA double-strand breaks. FA epithelial cells are highly sensitive to DNA cross-linking agents leading to genomic instability and increased risk for tumorigenesis. Oral cancer (OC) is 500-700 times higher in FA patients than the general population and occurs at a young age. FA OC patients cannot tolerate chemo-and radiation therapy; hence, surgery represents the best curative option if the OC can be diagnosed at early stage. Therefore, development of a minimally invasive diagnostic test for OC screening in FA patients is a critical and unmet need. In an ongoing clinical trial, we are using a programmable bio-nano-chip (p-BNC)-based assay to measure the cytomorphometry and biomarker levels of brush biopsies of potentially malignant oral mucosal. Data analyses revealed that a combination of molecular and cytomorphometric features distinguishes OC from non-neoplastic cells. Methods: In the current pilot study, we performed p-BNC-based assay to compare the cytomorphologic parameters and molecular biomarkers (αvβ6, EGFR, CD147, McM2, Geminin, and Ki67) in five different established OC cell lines from FA patients (OHSU-974, VU-1131, VU-1365) and non-FA patients (22A, 686Tu). In addition, we performed in vitro growth rate (MTT and BrdU-ELISA) and Matrigel invasion assays to characterize the phenotypic differences between these two groups of cell lines. Results: Preliminary morphological data from a Ki67 p-BNC assay indicated significant increases in the nuclear-cytoplasmic ratio, nuclear abnormalities and micronucleus formation in some of the FA-OC cell lines as compared to non-FA-OC cells (22A vs. VU-1131 and VU-1365, p=0.0010, p=0.0048, respectively; and 686Tu vs. VU-1365-T, p=0.0399). Comparison of molecular expression across biomarkers revealed that FA-OC cells expressed lower levels of McM2 (OHSU-974&gt;VU-1131&gt;VU-1365), Geminin and CD147 as compared to non-FA OC cells. Expression levels of αvβ6 were higher on VU-1365-T FA-OC cells than on non-FA-OC cells whereas expression of EGFR, and Ki67 was comparable across the two groups of cell lines. Non-FA-OC cells demonstrated significantly higher growth rate than FA-OC cells. On the other hand, FA-OC cells displayed significantly greater Matrigel invasiveness (mean # of Matrigel invasive cells/mean # of migrating cells x 100) than non-FA oral cancer cells. Conclusion: FA-OC cells exhibit more aggressive phenotype with unique cytomorphometric and biomarker-expression patterns compared to non-FA OC cells. Currently studies are in progress to evaluate the diagnostic capability of p-BNC system in analyzing non-invasive cytobrush specimens’ of potentially malignant oral mucosal lesions of FA patients.

Citation Format: Nadarajah Vigneswaran, Pierre Floriano, Jean Wu, Annie Wu, Ann Gillenwater, John McDevitt. Programmable bio-nano-chip (p-BNC) based cytomorphometric and biomarker expression analysis and phenotypic characterization of Fanconi anemia oral cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 55. doi:10.1158/1538-7445.AM2013-55