Abstract 4612: HOXA13 drives hepatocytes proliferation and liver tumorigenesis in mice

Abstract

Abstract Introduction: Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver and the third most common cause of cancer related mortality worldwide. For patients suffering from advanced stage disease, the few therapeutic options available are not curative and improve patient survival by only a few months. Therefore, new molecular targets that can be explored as therapeutic options are highly needed. Class I Homeobox (HOX) genes are fundamental components of embryonic patterning and morphogenesis, with expression persisting into adulthood. They are also implicated in neoplastic transformations. However, the role of HOX genes is poorly understood and the functional relationship between the malignant phenotype and abnormal expression of HOX genes is still unclear. In this study we sought to define the role of the HOXA13 gene in hepatocarcinogenesis using in vivo models. Methods: To unravel the molecular mechanism of HOXA13 driven tumorigenesis in liver and its direct oncogenicity in vivo, a murine model of HOXA13 overexpression in liver was generated using hydrodynamic injection coupled with a transposase system. This model led to the stable and specific HOXA13 expression in C57BL6\J mouse hepatocytes up to 5 months post injection. Mouse phenotype was followed over time, from 2 weeks up to 1 year post injection. 16 mice (8 for CTRL vector and 8 for HOXA13) were injected and sacrificed for every time point. RNA sequencing was performed to monitor the transcriptomic changes over time. Results: 1 year post injection 50% (4/8) of the injected mice with HOXA13 developed liver tumors of various histological grades and types, from very well differentiated HCCs to very highly undifferentiated and cholangiocarcinoma like nodules. HOXA13 overexpression in the liver led to highly proliferative hepatocytes after only 2 weeks and the proliferative phenotype was maintained until 5 months post injection, when pre neoplastic lesions began to form. HOXA13 overexpression correlated not only with proliferation but also with the DNA damage marker yH2AX, suggesting a possible mechanism of tumorigenesis driven by genome instability. Gene set enrichment analysis of RNA-seq performed on whole liver extracts of 2 week old mice and tumors showed that the main pathways involved in HOXA13 expression are cell cycle, in particular G2/M transition and mitotic assembly checkpoint, angiogenesis, TP53 pathway, IL6JAKSTAT3 signaling, Notch signaling and epithelial to mesenchymal transition. Conclusion: Our study highlights the key role of HOXA13 as a potential novel oncogene in HCC development and suggests possible mechanisms through which it drives liver tumorigenesis. We expect that the generated data in vivo, coupled with mass spectrometry and ChIP sequencing experiments performed in vitro, will further help us identify downstream effectors of HOXA13 thus providing new potential therapeutic targets for HCC. Citation Format: Gaia Bianco, Hesam Montazeri, Luca Quagliata, Tracy O'Connor, Ursula Ehmer, Rupert Oellinger, Mathias Matter, Christofori Gerhard M., Charlotte K.Y. Ng, Salvatore Piscuoglio, Mathias Heikenwaelder, Luigi M. Terracciano. HOXA13 drives hepatocytes proliferation and liver tumorigenesis in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4612.