Terminal differentiation is often coupled with permanent exit from the cell cycle and represents the most common cellular state in adult animals. Yet it remains unclear how proliferation is blocked in differentiated tissues. The goal of this project is to determine how terminal differentiation signals impinge on the cell cycle machinery to induce a stable quiescent state and investigate how this state is disrupted in cancer. In the research proposed here, I use a combination of genetic and biochemical approaches in Drosophila and mammalian cells to delineate the conserved genetic pathways that control cell cycle exit. During the K99 phase of this grant, I investigated the redundant mechanisms that limit cycling in differentiated Drosophila tissues, obtained experience working with 2 different mammalian cell lines that can be induced to differentiate in vitro for future studies of cell cycle exit, and worked on a screen to identify genes that when overexpressed or inhibited by RNAi, de-regulate the cell cycle to cause ectopic proliferation in contexts of terminal differentiation. Such genes would normally be activated or inhibited upon terminal differentiation, but likely become de-regulated in cancers. During the ROO phase of the award, I will determine the precise mechanisms by which these genes regulate the cell cycle (Aim 1) and examine the roles of mammalian orthologs of these genes in normal and cancerous mammalian cells (Aim2). I will also investigate the signaling pathways by which these genes are regulated, using genetic epistasis tools in both Drosophila and mammalian cells (Aim 3). Such work will begin to delineate the conserved pathways connecting terminal differentiation signals with cell cycle controls, and determine how they can become de-regulated leading to cancer.
Uncontrolled cell division is a hallmark of cancer. My research addresses how mature cells block the cell cycle to prevent inappropriate division. My proposed research will identify signals that when activated, give the wrong directions to mature cells and cause excessive division, mimicking cancer. By understanding how cell division can be improperly instructed, we hope to identify new targets for the treatment of cancer.
|Sun, Dan; Buttitta, Laura (2017) States of G0 and the proliferation-quiescence decision in cells, tissues and during development. Int J Dev Biol 61:357-366|
|Guo, Yongfeng; Flegel, Kerry; Kumar, Jayashree et al. (2016) Ecdysone signaling induces two phases of cell cycle exit in Drosophila cells. Biol Open 5:1648-1661|
|Bolin, Kelsey; Rachmaninoff, Nicholas; Moncada, Kea et al. (2016) miR-8 modulates cytoskeletal regulators to influence cell survival and epithelial organization in Drosophila wings. Dev Biol 412:83-98|
|Flegel, Kerry; Grushko, Olga; Bolin, Kelsey et al. (2016) Roles for the Histone Modifying and Exchange Complex NuA4 in Cell Cycle Progression in Drosophila melanogaster. Genetics 203:1265-81|
|O'Keefe, David D; Thomas, Sean; Edgar, Bruce A et al. (2014) Temporal regulation of Dpp signaling output in the Drosophila wing. Dev Dyn 243:818-32|
|Flegel, Kerry; Sun, Dan; Grushko, Olga et al. (2013) Live cell cycle analysis of Drosophila tissues using the Attune Acoustic Focusing Cytometer and Vybrant DyeCycle violet DNA stain. J Vis Exp :e50239|
|O'Keefe, David D; Thomas, Sean R; Bolin, Kelsey et al. (2012) Combinatorial control of temporal gene expression in the Drosophila wing by enhancers and core promoters. BMC Genomics 13:498|