Haploinsufficiency of CDK4 prevents hepatocarcinogenesis in mice deficient in ELF, a mediator of TGF-beta signaling

Abstract

10608 Background: The transforming growth factor-β (TGF-β) has a dual role in cancer, but in the prevention of tumorigenesis, TGF-β signaling inhibits cell cycle progression through an increase in cell cycle inhibitors and decrease in cell cycle activators. The adaptor protein embryonic liver fodrin (ELF) is crucial for normal TGF-β signaling. ELF facilitates the association and nuclear translocation of the TGF-β signaling proteins, Smad3 and Smad4. We have demonstrated that elf± mice develop hepatocellular cancers (HCC) spontaneously within 12 months. We have also shown that most human HCCs demonstrate a decreased expression of ELF. Our goal was to assess the interaction between ELF and cyclin dependent kinase 4 (CDK4), which is overexpressed in most human cancers. Methods: We compared the expression of CDK4 and ELF by immunohistochemistry and analyzed the interaction between CDK4 and ELF, in vitro by immunoprecitpitation and western blot. Finally, we assessed the development of HCC in intercrossed elf± and CDK4+/neo mice. Results: CDK4 and ELF expression are inversely correlated in HCCs from elf± mice. Furthermore, ELF interacts with CDK4 by forming a complex that includes Smad3 in COS-7 and HepG2 cells. ELF is phosphorylated upon overexpression of CDK4, revealing a possible mechanism by which CDK4 may inhibit ELF-dependent TGF-β signaling. To further assess whether the development of HCC in elf± mice is dependent upon CDK4 expression, we have intercrossed elf± and CDK4+/neo mice. At 12 months, only 17% of the elf±/CDK4+/neo mice developed HCC, compared to 40% of our historical control elf± mice. This initial data strongly suggests that haploinsufficiency of CDK4 can prevent the HCCs seen in elf± mice, and provides the genetic foundation for further exploring the benefits of specific inhibitors of CDK4 in the treatment of HCC. Conclusions: These results demonstrate the molecular interactions between ELF and CDK4, and suggest a mechanism by which CDK4 may render cells unresponsive to the growth inhibitory effects of TGF-β signaling. Furthermore, haploinsufficiency of CDK4 prevents the formation of HCC, and thus targeted-inhibition of CDK4 activity may be a logical treatment for HCC in humans. No significant financial relationships to disclose.