Accurate chromosome segregation is essential for the propagation of species and the viability of cells, and is driven by a complex microtubule-based structure called the spindle. Intensive biochemical, genetic, and proteomic efforts provide an extensive catalogue of proteins that participate in spindle organization and spindle-dependent chromosome movement. However, these efforts don't reveal the molecular mechanisms that ensure faithful chromosome segregation in mammalian cells. Recently, we showed that the most common cause of chromosome mis-segregation in human tumor cells is the persistence of kinetochore-microtubule (K- MT) attachment errors. However, our understanding for how K-MT attachments are regulated to promote error correction remains starkly incomplete. We don't understand how different molecular components create a coherent output to fine-tune K-MT attachment stability during cell cycle transitions. We also don't understand how aneuploidy influences genome stability and cell survival. It is our goal in the forthcoming funding period to combine biochemical methods and live cell imaging to test models and determine the mechanisms of the regulation of K-MT attachments in mitosis. Understanding these mechanisms could lead to new therapeutic approaches for cancer treatment.
The specific aims of this proposal are, 1.) To use live cell imaging to determine the biochemical changes underlying the transition from prometaphase to metaphase;2.) To combine live cell imaging with molecular tools to test the model that changes in the copy number of single genes is sufficient to undermine faithful chromosome segregation by disrupting K-MT attachment dynamics;3.) To quantify the genetic damage caused by persistent K-MT attachment errors;and 4.) To test mechanisms that suppress aneuploid cell growth in live animals.

Public Health Relevance

Errors in chromosome segregation cause aneuploidy that causes birth defects and is commonly associated with advanced stage cancer. The goal of the experiments proposed here is to combine biochemical methods and live cell imaging to identify the proteins and determine the mechanisms responsible for high fidelity chromosome segregation in human cells. Data generated from this work will provide insight into mechanisms of aneuploidy in tumor cells and may reveal strategies for therapy of chromosomally unstable aneuploid tumors.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
3R37GM051542-18S1
Application #
8901358
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Deatherage, James F
Project Start
1996-08-01
Project End
2018-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
18
Fiscal Year
2014
Total Cost
$64,800
Indirect Cost
$24,800
Name
Dartmouth College
Department
Biochemistry
Type
Schools of Medicine
DUNS #
041027822
City
Hanover
State
NH
Country
United States
Zip Code
03755
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Orr, Bernardo; Talje, Lama; Liu, Zhexian et al. (2016) Adaptive Resistance to an Inhibitor of Chromosomal Instability in Human Cancer Cells. Cell Rep 17:1755-1763
Kim, Jung-Sik; He, Xiaoyuan; Orr, Bernardo et al. (2016) Intact Cohesion, Anaphase, and Chromosome Segregation in Human Cells Harboring Tumor-Derived Mutations in STAG2. PLoS Genet 12:e1005865
Orr, Bernardo; Godek, Kristina M; Compton, Duane (2015) Aneuploidy. Curr Biol 25:R538-42
Hu, Shanhu; Danilov, Alexey V; Godek, Kristina et al. (2015) CDK2 Inhibition Causes Anaphase Catastrophe in Lung Cancer through the Centrosomal Protein CP110. Cancer Res 75:2029-38
Meppelink, Amanda; Kabeche, Lilian; Vromans, Martijn J M et al. (2015) Shugoshin-1 balances Aurora B kinase activity via PP2A to promote chromosome bi-orientation. Cell Rep 11:508-15
Bakhoum, Samuel F; Kabeche, Lilian; Wood, Matthew D et al. (2015) Numerical chromosomal instability mediates susceptibility to radiation treatment. Nat Commun 6:5990
Godek, Kristina M; Kabeche, Lilian; Compton, Duane A (2015) Regulation of kinetochore-microtubule attachments through homeostatic control during mitosis. Nat Rev Mol Cell Biol 16:57-64
Hu, Shanhu; Lu, Yun; Orr, Bernardo et al. (2015) Specific CP110 Phosphorylation Sites Mediate Anaphase Catastrophe after CDK2 Inhibition: Evidence for Cooperation with USP33 Knockdown. Mol Cancer Ther 14:2576-85
Bakhoum, Samuel F; Kabeche, Lilian; Murnane, John P et al. (2014) DNA-damage response during mitosis induces whole-chromosome missegregation. Cancer Discov 4:1281-9

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