Abstract 502: Development of lung cancer cell lines exhibiting cancer stem cell properties through application of stem-cell culturing techniques
Authors: Peter Young, Penny Roberts, Xiaolin Xu, Jeff Hooley, Doug Smith, Paul A. Moore, Jennie Mather
Journal: Cancer Research (2011)

Abstract Introduction: Lung cancer is the most common cause of cancer death worldwide: with greater than 160,000 deaths reported in the US in 2008 and a dismal 15% 5-year survival rate, there is a clear need for improved therapeutic approaches. Considering the increasing evidence for the role of cancer stem cells (CSC) in driving tumor initiation and metastases, development of model systems to isolate and characterize such populations in lung cancer would provide important tools to drive discovery of novel therapeutic modalities. We recently reported application of selective culture conditions to generate novel cell lines from colon cancer that exhibit CSC properties (1). Here we apply a similar approach to lung cancer. Methodology: Freshly resected NSLC tissue was obtained, processed and cultured using serum-free defined culture media previously developed for fetal lung stem cells (2). Expanded cell lines were characterized for their tumor initiating properties in NSG mice upon implantation under the sub-renal capsule. Cell populations generated from lung cancer under serum-containing conditions served as controls. Tumors were analyzed by H&amp;E and immunostaining for specific markers. Gene expression profiles were determined by U133 Affymetrix arraying, and the cell surface protein profile examined by flow cytometry using a bank of approximately ∼200 cell surface reactive mAbs. Results: Lung cancer cell lines, were successfully cultured from independent lung tumor specimens and remained genetically stable upon prolonged passaging in vitro. One such line, LUCA22, was derived from an adenosquamous carcinoma specimen and harbors a KRAS mutation. This line formed tumors in mice within 10 weeks upon implantation of as few as 500 cells; these tumors recapitulated the morphology of the patient's original tumor, with mixed features that included differentiation into both adeno- and squamous-type carcinoma. Within 16 weeks, spontaneous metastases were observed to the lung, liver and pancreas. Gene array analyses of LUCA22 cells compared to control populations revealed up-regulation of genes associated with the Wnt and TGF-beta pathways, while flow cytometry identified over-expression of several CSC-associated cell surface proteins, including CD44. Conclusion: Novel NSCL cell lines have been derived from primary tumor specimens by employing culture conditions previously established for tissue stem cells. These lines display key features ascribed to CSC including the capacity to recapitulate the original tumor morphology from where they were derived and the ability to spontaneously metastasize. Gene array analysis and mAbs library panning of the cell lines are being applied to identify lead opportunities for targeting lung cancer stem cells. References: (1) Roberts et al., 2010, ISSCR Annual Meeting Abstract #840; (2) Roberts et al., 1990, Amer J Physiol: 3:415 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 502. doi:10.1158/1538-7445.AM2011-502