Abstract 5028: Reverse translational research for identifying the correlation between EGFR mutation status and DPD protein expression in lung adenocarcinoma
Authors: Tsuchiya Tomoshi , Tominaga Tetsuo , Mochinaga Koji , Yamasaki Naoya , Matsumoto Keitaro , Miyazaki Takuro , Hatachi Go , Kitajima Yuka , Nagayasu Takeshi 
Published: July 2016

[Background] Reverse translation research is often described as “From the Bedside to the Bench”, which means taking an understanding gained by observations in a patient population to create hypotheses and confirmation of the hypothesis in the laboratory. According to current chemotherapy treatment for lung adenocarcinoma, empirically, clinicians know that epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKI) tend to be effective for radiological ground glass nodule- type lung adenocarcinomas. On the other hand, it has been shown that 5-FU sensitivity in patients with non-small cell lung cancer (NSCLC) is associated with EGFR mutation status. From such clinical hints, we identified a relationship between dihydropyrimidine dehydrogenase (DPD), a 5-FU degrading enzyme, and EGFR mutation status.

[Methods] We firstly focused on clinicopathologic factors and in vitro correlations between DPD expression and EGFR mutation status. Secondly, we analyzed crosstalk between the EGFR signal cascade and DPD protein expression using EGFR mutated and not mutated cell lines.

[Clinicopathological Study] EGFR mutations and messenger RNA (mRNA) levels of DPD were analyzed in 47 resected NSCLC tumors by laser-capture microdissection. EGFR mutation status and the immunohistochemical expression of DPD in 49 resected NSCLC tumors were also examined. As a result, adenocarcinoma in situ showed significantly higher DPD mRNA levels and more EGFR mutation frequency than other histological types (P &lt; 0.05). DPD immunopositive cases were more frequently observed in adenocarcinoma, in females, and in nonsmokers; which were correlated with EGFR mutation status (P &lt; 0.003).

[Basic Study] In vitro, EGFR-mutated cell lines (PC9, HCC827; exon19 E746-A750 and H1975; exon21 L858R, T790M, gefitinib resistant) showed higher DPD mRNA and protein expression than wild types (H1437, H1299). The DPD gene (DPYD), and DPD protein expression were known to be regulated by the transcription factor Sp1. In the PC9 study, EGF treatment induced up-regulation of both Sp1 and DPD. Gefitinib, an EGFR-TKI, and mithramycin A, specific Sp-1 inhibitors, suppressed them. However, the phenomena were not observed in EGFR-wild types and EGFR-TKI resistant cell lines. These results suggest DPD is regulated mainly in EGFR signal cascade enhanced lung cancers such as EGFR mutated adenocarcinomas. 5-FU and its derivatives such as uracil-tegafur (UFT) or S-1 might be effective for EGFR wild type adenocarcinomas because of the low DPD expression.

[Conclusion] Reverse translational research can provide useful information for clinical treatment. In the present study, we identified crosstalk between the EGFR signal cascade and DPD protein expression, which might affect therapeutic strategy for lung adenocarcinomas.

Citation Format: Tomoshi Tsuchiya, Tetsuo Tominaga, Koji Mochinaga, Naoya Yamasaki, Keitaro Matsumoto, Takuro Miyazaki, Go Hatachi, Yuka Kitajima, Takeshi Nagayasu. Reverse translational research for identifying the correlation between EGFR mutation status and DPD protein expression in lung adenocarcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5028.