Abstract B02: Protein Kinase C iota is required for the maintenance of a tumor-initiating cell phenotype in lung squamous cell carcinoma.

Abstract

Abstract Background: Protein kinase C iota (PKCι) is an oncogene that is frequently targeted for tumor-specific gene amplification in human cancers harboring the 3q26 amplicon including Non-Small Cell Lung Carcinoma (NSCLC). PKCι is required for the maintenance of the transformed phenotype of NSCLC cells in vitro and in vivo. In addition, PKCι is critical for lung tumorigenesis in the LSL-K-rasG12D mouse model of NSCLC. Recently, we demonstrated that genetic disruption of Prkcι/λ; in LSL-K-rasG12D mice blocks K-ras-mediated expansion and morphological transformation of bronchioalveolar stem cells (BASCs), the first identifiable step in K-ras-mediated transformation in this model. These results demonstrate that PKCι plays a requisite role in tumor initiation, and suggest that PKCι may serve a similar role in the tumor-initiating activity of lung cancer stem cells. Our current studies aim to identify and characterize the molecular signaling pathway(s) by which PKCι regulates the biology of human lung squamous cell carcinoma (LSCC) tumor initiating cells (TICs) isolated from LSCC cell lines and primary tumors. Methods: Cultures enriched in TICs were established from LSCC lines or primary human LSCC tumors that harbor PRKCI amplification. PKCι expression in TIC cultures was inhibited by lentiviral shRNA, and the effect of PKCι knockdown on their stem-like properties was assessed. Whole genome deep sequencing analysis was performed on mRNA from parental, control TIC and PKCι knockdown TIC cultures in order to identify PKCι-regulated targets and signaling pathways in TICs. Results: LSCC TIC cultures express putative stem cell markers, and exhibit clonal expansion, self-renewal, enhanced anchorage-independent growth and tumorigenic potential in vivo. RNAi-mediated knockdown of PKCι inhibited TIC growth, clonal expansion, transformed growth and tumor initiation. mRNA sequencing revealed that PKCι regulates the expression and function of critical molecules involved in the stem-like phenotype of LSCC TICs. Details of PKCι-mediated signaling in LSCC TICs will be discussed. Conclusions: PKCι is required for maintenance of a highly tumorigenic stem-like population of cells in human LSCC. Given the prevalence of 3q26 amplification in human cancer, PKCι may regulate the biology of TICs in tumor types other than LSCC. Our results provide a rationale for therapeutic targeting of PKCι in LSCC TICs. Citation Format: Verline Justilien, Michael P. Walsh, Syed A. Ali, E. A. Thompson, Alan P. Fields. Protein Kinase C iota is required for the maintenance of a tumor-initiating cell phenotype in lung squamous cell carcinoma. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr B02.