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CDK2 inhibition disorders centrosome stoichiometry and alters cellular outcomes in aneuploid cancer cells
Zibo Chen1, Xi Liu1, Masanori Kawakami1
1Molecular Pharmacology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
Abstract:
Cyclin-dependent Kinase 2 (CDK2) inhibition prevents supernumerary centrosome clustering. This causes multipolarity, anaphase catastrophe and apoptotic death of aneuploid cancers. This study elucidated how CDK2 antagonism affected centrosome stoichiometry. Focused ion beam scanning electron microscopy (FIB-SEM) and immunofluorescent imaging were used. Studies interrogated multipolar mitosis after pharmacologic or genetic repression of CDK2. CDK2/9 antagonism with CYC065 (Fadraciclib)-treatment disordered centrosome stoichiometry in aneuploid cancer cells, preventing centrosome clustering. This caused ring-like chromosomes or multipolar cancer cells to form before onset of cell death. Intriguingly, CDK2 inhibition caused a statistically significant increase in single centrioles rather than intact centrosomes with two centrioles in cancer cells having chromosome rings or multipolarity. Statistically significant alterations in centrosome stoichiometry were undetected in other mitotic cancer cells. To confirm this pharmacodynamic effect, CDK2 but not CDK9 siRNA-mediated knockdown augmented cancer cells with chromosome ring or multipolarity formation. Notably, engineered gain of CDK2, but not CDK9 expression, reversed emergence of cancer cells with chromosome rings or multipolarity, despite CYC065-treatment. In marked contrast, CDK2 inhibition of primary human alveolar epithelial cells did not confer statistically significant increases of cells with ring-like chromosomes or multipolarity. Hence, CDK2 antagonism caused differential effects in malignant versus normal alveolar epithelial cells. Translational relevance was confirmed by CYC065-treatment of syngeneic lung cancers in mice. Mitotic figures in tumors exhibited chromosome rings or multipolarity. Thus, CDK2 inhibition preferentially disorders centrosome stoichiometry in cancer cells. Engaging this disruption is a strategy to explore against aneuploid cancers in future clinical trials.
Insights
Inhibiting Cyclin-dependent Kinase 2 (CDK2) disrupts centrosome balance in aneuploid cancer cells, leading to cell death. This targeted approach shows promise for treating cancers with abnormal chromosome numbers.
Area of Science:
- Cell Biology
- Cancer Biology
- Molecular Oncology
Background:
- Aneuploid cancers often exhibit supernumerary centrosomes, leading to multipolar mitosis and genomic instability.
- Cyclin-dependent Kinase 2 (CDK2) plays a role in regulating centrosome duplication and segregation.
Purpose of the Study:
- To investigate the effect of CDK2 antagonism on centrosome stoichiometry in aneuploid cancer cells.
- To determine if CDK2 inhibition can prevent supernumerary centrosome clustering and induce cancer cell death.
Main Methods:
- Focused ion beam scanning electron microscopy (FIB-SEM) and immunofluorescent imaging were employed.
- Pharmacologic (CYC065) and genetic (siRNA, gene expression) repression of CDK2 were utilized.
- Experiments were conducted in cancer cell lines, primary human alveolar epithelial cells, and syngeneic mouse lung cancer models.
Main Results:
- CDK2 antagonism with CYC065 disordered centrosome stoichiometry, preventing clustering and causing multipolarity or ring-like chromosomes in aneuploid cancer cells.
- CDK2 inhibition significantly increased single centrioles in affected cancer cells.
- CDK2 knockdown mimicked these effects, while CDK2 overexpression reversed them, demonstrating specificity.
- CDK2 inhibition did not cause significant multipolarity in normal alveolar epithelial cells, indicating differential effects.
Conclusions:
- CDK2 antagonism preferentially disorders centrosome stoichiometry in cancer cells, leading to mitotic catastrophe and cell death.
- This disruption of centrosome balance presents a potential therapeutic strategy for aneuploid cancers.
- Targeting CDK2 offers a selective approach to combatting cancer with potential for clinical translation.
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