Our overall goal is to understand the mechanisms by which dividing cancer and normal cells are killed, or escape death, when treated with diverse anti-mitotic drugs. We seek understanding both at the level of cell population behaviors, and the molecular mechanisms that give rise to those behaviors. We are especially interested in understanding how different classes of anti-mitotic drug differ in their ability to kill cancer cells, and using that information to design better future drugs. We will address these goals in 4 specific aims:
Aim 1. How do drugs with different anti-mitotic mechanisms differ in their ability to kill cancer cells at the level of cell population behavior? We will use microscopy with fluorescent reporters for key molecular events to measure single-cell responses to four anti-mitotic drugs across a panel of cell lines chosen to vary in apoptosis sensitivity. We will also test the effects of blocking different aspects ofthe drug response.
Aim 2. Elucidate the molecular mechanism of cell death during mitotic arrest. To determine how the intrinsic apoptosis pathway is activated by prolonged mitotic arrest we will test candidate regulators of apoptosis, and pursue an unbiased biochemical approach. We will also Identify an alternative cell death pathway cells use when they cannot escape mitotic arrest, and apoptosis is blocked.
Aim 3. Develop small molecule inhibitors of mitotic exit that work independent of the SAC. Our preliminary data suggest that novel anti-mitotic drugs with this mechanism would kill cancer cells more effectively than current drugs. We will test this concept, and identify druggable targets in the mitotic exit pathway, by small molecule screening in Core B using a cell-based assay, followed by identification of protein targets of hits.
Aim 4. Compare drug responses in mouse tumors to those seen in cell culture. Drug response mechanisms elucidated in cell culture in aims 1-3 may not translate to real tumors. We will test whether cells that die during mitotic arrest in mouse tumors cause bystander killing of non-dividing tumor cells, which could explain why some tumors with low mitotic index can be treated with anti-mitotic drugs. Working with Core C we will probe responses to anti-mitotic drugs in mouse tumors at the single-cell level by intravital imaging.
Chemotherapy drugs that block division of cancer cells are important in cancer treatment, but we need better methods for predicting which patients will respond well. We also need new drugs that are more effective at killing dividing cancer cells. We will address these challenges by discovering how cancer cells are killed by current drugs, why some cancer cells escape killing, and by testing a concept for a better future drug.
|Sharma, Mayuri; Kamil, Jeremy P; Coughlin, Margaret et al. (2014) Human cytomegalovirus UL50 and UL53 recruit viral protein kinase UL97, not protein kinase C, for disruption of nuclear lamina and nuclear egress in infected cells. J Virol 88:249-62|
|Laughney, Ashley M; Kim, Eunha; Sprachman, Melissa M et al. (2014) Single-cell pharmacokinetic imaging reveals a therapeutic strategy to overcome drug resistance to the microtubule inhibitor eribulin. Sci Transl Med 6:261ra152|
|Kim, Eunha; Yang, Katherine S; Giedt, Randy J et al. (2014) Red Si-rhodamine drug conjugates enable imaging in GFP cells. Chem Commun (Camb) 50:4504-7|
|Lee, Sungon; Vinegoni, Claudio; Sebas, Matthew et al. (2014) Automated motion artifact removal for intravital microscopy, without a priori information. Sci Rep 4:4507|
|Turetsky, Anna; Kim, Eunha; Kohler, Rainer H et al. (2014) Single cell imaging of Bruton's tyrosine kinase using an irreversible inhibitor. Sci Rep 4:4782|
|Lee, Robin E C; Walker, Sarah R; Savery, Kate et al. (2014) Fold change of nuclear NF-?B determines TNF-induced transcription in single cells. Mol Cell 53:867-79|
|Pan, Rongqing; Hogdal, Leah J; Benito, Juliana M et al. (2014) Selective BCL-2 inhibition by ABT-199 causes on-target cell death in acute myeloid leukemia. Cancer Discov 4:362-75|
|Foijer, Floris; Xie, Stephanie Z; Simon, Judith E et al. (2014) Chromosome instability induced by Mps1 and p53 mutation generates aggressive lymphomas exhibiting aneuploidy-induced stress. Proc Natl Acad Sci U S A 111:13427-32|
|Krukenberg, Kristin A; Jiang, Ruomu; Steen, Judith A et al. (2014) Basal activity of a PARP1-NuA4 complex varies dramatically across cancer cell lines. Cell Rep 8:1808-18|
|Xia, X; Owen, M S; Lee, R E C et al. (2014) Cell-to-cell variability in cell death: can systems biology help us make sense of it all? Cell Death Dis 5:e1261|
Showing the most recent 10 out of 37 publications