Many human tumor cells are chromosomally unstable, showing an elevated rate of gains and losses of whole chromosomes that is 10-100 times higher than that seen in diploid primary cells. Chromosome instability (CIN) enhances the evolution of tumor cells causing genomic changes that can promote metastasis and chemotherapeutic resistance. CIN can have a causal role in tumorigenesis and it correlates with poor patient prognosis. Identifying the mutations that cause CIN in human cancer, and understanding why they reduce the fidelity of mitosis, are important objectives in cancer research. The inactivation of the pRB pathway promotes cell proliferation and is a common event in tumor cells. Interestingly, the functional inactivation of pRB causes aneuploidy. Recent experiments show that the specific loss of pRB increases rates of chromosome mis-segregation to levels that are remarkably similar to CIN tumor cells. This strongly suggests that a significant fraction of the chromosome instability seen in tumor cells is a byproduct of the inactivation of pRB. This grant investigates the exciting link between pRB inactivation and chromosome mis-segregation. Newly obtained results show that the loss of pRB causes defects in centromere function and chromatid cohesion. We will test the hypothesis that defects in cohesion and condensation allow chromosome mis-segregation when pRB-deficient cells are delayed in mitosis. Cohesin is the primary determinant of chromosome cohesion.
In Aim 1 we will determine how the association of cohesin complexes with chromatin is altered in the absence pRB, and will test the functional significance of physical interactions between pRB and cohesin. The experiments in Aim 2 will test whether the mitotic defects in pRB-deficient cells can be either suppressed or enhanced. Such experiments may lead to new therapeutic opportunities.
In Aim 3 we will extend these studies to cancer cells and will measure and compare the changes in centromere function, chromosome cohesion, and the rates of chromosome mis-segregation seen when functional pRB is re-introduced into tumor cells that lack it, or is specifically removed from tumor cells with functional pRB. We will determine whether similar changes occur when pRB is inactivated by deregulated cdk activity and when pRB-related proteins are targeted.
Cancer is one of the leading causes of mortality in the United States and this proposal investigates the molecular changes that cause cancer cells to mis-segregate chromosomes. Chromosome instability helps tumors to evolve, promoting metastasis and chemotherapeutic resistance. Understanding how the inactivation of RB reduces the fidelity of mitosis may reveal ways to suppress chromosome instability.
|Manning, Amity L; Yazinski, Stephanie A; Nicolay, Brandon et al. (2014) Suppression of genome instability in pRB-deficient cells by enhancement of chromosome cohesion. Mol Cell 53:993-1004|
|Heilmann, Andreas M; Perera, Rushika M; Ecker, Veronika et al. (2014) CDK4/6 and IGF1 receptor inhibitors synergize to suppress the growth of p16INK4A-deficient pancreatic cancers. Cancer Res 74:3947-58|
|Wang, Bin; Kettenbach, Arminja N; Gerber, Scott A et al. (2014) Neurospora WC-1 recruits SWI/SNF to remodel frequency and initiate a circadian cycle. PLoS Genet 10:e1004599|
|Manning, A L; Benes, C; Dyson, N J (2014) Whole chromosome instability resulting from the synergistic effects of pRB and p53 inactivation. Oncogene 33:2487-94|
|Black, Joshua C; Manning, Amity L; Van Rechem, Capucine et al. (2013) KDM4A lysine demethylase induces site-specific copy gain and rereplication of regions amplified in tumors. Cell 154:541-55|
|Evertts, Adam G; Manning, Amity L; Wang, Xin et al. (2013) H4K20 methylation regulates quiescence and chromatin compaction. Mol Biol Cell 24:3025-37|
|Zhang, Jinghui; Benavente, Claudia A; McEvoy, Justina et al. (2012) A novel retinoblastoma therapy from genomic and epigenetic analyses. Nature 481:329-34|
|Manning, Amity L; Dyson, Nicholas J (2012) RB: mitotic implications of a tumour suppressor. Nat Rev Cancer 12:220-6|