Identification of Regulators of Polyploidization Presents Therapeutic Targets for Treatment of AMKL

Division of Hematology/Oncology, Northwestern University, Chicago, IL 60611, USA.
Cell (Impact Factor: 32.24). 08/2012; 150(3):575-89. DOI: 10.1016/j.cell.2012.06.032
Source: PubMed


The mechanism by which cells decide to skip mitosis to become polyploid is largely undefined. Here we used a high-content image-based screen to identify small-molecule probes that induce polyploidization of megakaryocytic leukemia cells and serve as perturbagens to help understand this process. Our study implicates five networks of kinases that regulate the switch to polyploidy. Moreover, we find that dimethylfasudil (diMF, H-1152P) selectively increased polyploidization, mature cell-surface marker expression, and apoptosis of malignant megakaryocytes. An integrated target identification approach employing proteomic and shRNA screening revealed that a major target of diMF is Aurora kinase A (AURKA). We further find that MLN8237 (Alisertib), a selective inhibitor of AURKA, induced polyploidization and expression of mature megakaryocyte markers in acute megakaryocytic leukemia (AMKL) blasts and displayed potent anti-AMKL activity in vivo. Our findings provide a rationale to support clinical trials of MLN8237 and other inducers of polyploidization and differentiation in AMKL.

Download full-text


Available from: Ling-Zhi Wang
  • Source
    • "In contrast, MLN8237 caused an initial increase in S10 phosphorylation at lower concentrations, followed by a sharp drop at higher concentrations (Figures 1D; Figures S1E and S1F). This increase in phosphorylation of histone H3 in response to MLN8237 has been described previously and results from Aurora A inhibition with feedback increase in Aurora B activity (Gö rgü n et al., 2010; Wen et al., 2012). CD532 behaves similarly to MLN8237 with regard to histone H3 phosphorylation, consistent with an Aurora-A-selective effect. "
    [Show abstract] [Hide abstract]
    ABSTRACT: MYC proteins are major drivers of cancer yet are considered undruggable because their DNA binding domains are composed of two extended alpha helices with no apparent surfaces for small-molecule binding. Proteolytic degradation of MYCN protein is regulated in part by a kinase-independent function of Aurora A. We describe a class of inhibitors that disrupts the native conformation of Aurora A and drives the degradation of MYCN protein across MYCN-driven cancers. Comparison of cocrystal structures with structure-activity relationships across multiple inhibitors and chemotypes, coupled with mechanistic studies and biochemical assays, delineates an Aurora A conformation-specific effect on proteolytic degradation of MYCN, rather than simple nanomolar-level inhibition of Aurora A kinase activity.
    Full-text · Article · Aug 2014 · Cancer Cell
  • Source
    • "Preclinical testing has demonstrated preferential sensitivity of AML cell lines and primary AML specimens to Aurora A and Aurora B kinase inhibitors (91, 92). These agents may have particular therapeutic relevance for patients with acute megakaryoblastic leukemia (AMKL) based upon preclinical data demonstrating increased megakaryocyte polyploidization and AMKL cytotoxicity in vitro and in vivo with the Aurora kinase A inhibitor alisertib (MLN8237) (93). Clinical trials are in progress to evaluate the safety and/or efficacy of Aurora kinase inhibition as monotherapy or in conjunction with chemotherapy in adults with AML (NCT01779843) (33). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Approximately two-thirds of children with acute myeloid leukemia (AML) are cured with intensive multi-agent chemotherapy. However, refractory and relapsed AML remains a significant source of childhood cancer mortality, highlighting the need for new therapies. Further therapy intensification with traditional cytotoxic chemotherapy in pediatric AML is not feasible given the risks of both short-term and long-term organ dysfunction. Substantial emphasis has been placed upon the development of molecularly targeted therapeutic approaches for adults and children with high-risk subtypes of AML with the goal of improving remission induction and minimizing relapse. Several promising agents are currently in clinical testing or late preclinical development for AML, including monoclonal antibodies against leukemia cell surface proteins, kinase inhibitors, proteasome inhibitors, epigenetic agents, and chimeric antigen receptor engineered T cell immunotherapies. Many of these therapies have been specifically tested in children with relapsed/refractory AML in Phase 1 and 2 trials with a smaller number of new agents under Phase 3 evaluation for children with de novo AML. Although successful identification and implementation of new drugs for children with AML remain a formidable challenge, enthusiasm for novel molecular therapeutic approaches is great given the potential for significant clinical benefit for children who do not have other curative options.
    Full-text · Article · Mar 2014 · Frontiers in Oncology
  • Source
    • "A recent study indicated that a higher concentration (>5 μM) of H-1152 inhibited Aurora-A activity and induced polyploidization in acute megakaryocytic leukemia cells. In this study, 1 μM of H-1152 was used to induce centrosome fragmentation, in which Aurora-A activity was not affected [31]. The same study claimed that H-1152 is well tolerated by mice carrying a xenograft. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Accurate chromosome segregation is vital for cell viability. Many cancer cells show chromosome instability (CIN) due to aberrant expression of the genes involved in chromosome segregation. The induction of massive chromosome segregation errors in such cancer cells by small molecule inhibitors is an emerging strategy to kill these cells selectively. Here we screened and characterized small molecule inhibitors which cause mitotic chromosome segregation errors to target cancer cell growth. We screened about 300 chemicals with known targets, and found that Rho-associated coiled-coil kinase (ROCK) inhibitors bypassed the spindle assembly checkpoint (SAC), which delays anaphase onset until proper kinetochore-microtubule interactions are established. We investigated how ROCK inhibitors affect chromosome segregation, and found that they induced microtubule-dependent centrosome fragmentation. Knockdown of ROCK1 and ROCK2 revealed their additive roles in centrosome integrity. Pharmacological inhibition of LIMK also induced centrosome fragmentation similar to that by ROCK inhibitors. Inhibition of ROCK or LIMK hyper-stabilized mitotic spindles and impaired Aurora-A activation. These results suggested that ROCK and LIMK are directly or indirectly involved in microtubule dynamics and activation of Aurora-A. Furthermore, inhibition of ROCK or LIMK suppressed T cell leukemia growth in vitro, but not peripheral blood mononuclear cells. They induced centrosome fragmentation and apoptosis in T cell leukemia cells. These results suggested that ROCK and LIMK can be a potential target for anti-cancer drugs.
    Full-text · Article · Mar 2014 · PLoS ONE
Show more