VISAGE Reveals a Targetable Mitotic Spindle Vulnerability in Cancer Cells

Patterson, Jesse C.; Joughin, Brian A.; Prota, A.E.; Mühlethaler, Tobias; Jonas, Oliver; Whitman, Matthew A.; Varmeh, Shohreh; Chen, Sen; Balk, Steven P.; Steinmetz, Michel O.; Lauffenburger, Douglas A.; Yaffe, Michael B.

doi:10.1016/j.cels.2019.05.009

Cited by 24 publications

(44 citation statements)

References 92 publications

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“…The two compounds have indistinguishable chemical-genetic interactions with genes involved in modulating the microtubule network (KIF2C and TACC3), both destabilize microtubules in cells and in vitro, and both show substantially reduced toxicity in cell lines expressing a rationally-designed mutant of tubulin (L240F TUBB mutant), in which the rigosertib binding site in tubulin is mutated. Importantly, the specificity of the L240F TUBB mutant for microtubule-destabilizing agents, which is disputed by Reddy and co-workers, was recently confirmed by an independent research group (Patterson et al, 2019). We conclude that rigosertib kills cancer cells by destabilizing microtubules, in agreement with our original findings.…”

supporting

confidence: 90%

“…They found that TH588 synergizes with PLK1 inhibition in a manner that is independent of inhibition of MTH1, and through in-depth analysis found that TH588 destabilizes microtubules by binding in the same site as rigosertib. The L240F TUBB mutant indeed conferred resistance to TH588, but not to BI2536, which they used as a control (Patterson et al, 2019). Although this observation is published, Reddy and co-workers do not address the conflicting results, which are in support of our conclusions.…”

Section: Specificity Of Resistance Conferred By the L240f Tubb Mutantmentioning

confidence: 52%

“…Although these contradictions are difficult to reconcile without detailed knowledge of the protocols used to conduct their experiments, as discussed below our assays were designed to consistently provide higher specificity to detect the phenotypes in question. In addition, a key claim by Reddy and coworkers regarding the specificity of the rationally designed rigosertib-resistant TUBB mutant is contradicted by results published by independent investigators (Patterson et al, 2019).…”

Section: Discussionmentioning

confidence: 96%

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Pharmaceutical-grade Rigosertib is a Microtubule-destabilizing Agent

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Chen

Gilbert

et al. 2020

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We recently used CRISPRi/a-based chemical-genetic screens and targeted cell biological, biochemical, and structural assays to determine that rigosertib, an anti-cancer agent in phase III clinical trials, kills cancer cells by destabilizing microtubules. In a recent manuscript, Reddy and co-workers suggest that this microtubule-destabilizing activity of rigosertib is mediated not by rigosertib itself but by a contaminating degradation product of rigosertib, ON01500, present in formulations obtained from commercial vendors (Baker et al., 2019). Here, we demonstrate that treatment of cells with pharmaceutical-grade rigosertib (>99.9% purity) results in qualitatively indistinguishable phenotypes as treatment with commercially obtained rigosertib across multiple assays. The two compounds have indistinguishable chemical-genetic interactions with genes involved in modulating the microtubule network (KIF2C and TACC3), both destabilize microtubules in cells and in vitro, and both show substantially reduced toxicity in cell lines expressing a rationally-designed mutant of tubulin (L240F TUBB mutant), in which the rigosertib binding site in tubulin is mutated. Importantly, the specificity of the L240F TUBB mutant for microtubule-destabilizing agents, which is disputed by Reddy and co-workers, was recently confirmed by an independent research group (Patterson et al., 2019). We conclude that rigosertib kills cancer cells by destabilizing microtubules, in agreement with our original findings.

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supporting

confidence: 90%

Section: Specificity Of Resistance Conferred By the L240f Tubb Mutantmentioning

confidence: 52%

Section: Discussionmentioning

confidence: 96%

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Pharmaceutical-grade Rigosertib is a Microtubule-destabilizing Agent

Jost

Chen

Gilbert

et al. 2020

Preprint

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“…Emerging evidence connect mitotic arrest with ROS. For instance, it has been shown that the level of ROS changes with cell cycle, where ROS is increased during mitosis and a mitotic arrest further enhances ROS levels [41]. Whether MTH1 directly alter ROS levels and plays a role during mitosis is still under debate.…”

Section: Discussionmentioning

confidence: 99%

AXL and CAV-1 play a role for MTH1 inhibitor TH1579 sensitivity in cutaneous malignant melanoma

et al. 2020

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Cutaneous malignant melanoma (CMM) is the deadliest form of skin cancer and clinically challenging due to its propensity to develop therapy resistance. Reactive oxygen species (ROS) can induce DNA damage and play a significant role in CMM. MTH1 protein protects from ROS damage and is often overexpressed in different cancer types including CMM. Herein, we report that MTH1 inhibitor TH1579 induced ROS levels, increased DNA damage responses, caused mitotic arrest and suppressed CMM proliferation leading to cell death both in vitro and in an in vivo xenograft CMM zebrafish disease model. TH1579 was more potent in abrogating cell proliferation and inducing cell death in a heterogeneous coculture setting when compared with CMM standard treatments, vemurafenib or trametinib, showing its broad anticancer activity. Silencing MTH1 alone exhibited similar cytotoxic effects with concomitant induction of mitotic arrest and ROS induction culminating in cell death in most CMM cell lines tested, further emphasizing the importance of MTH1 in CMM cells. Furthermore, overexpression of receptor tyrosine kinase AXL, previously demonstrated to contribute to BRAF inhibitor resistance, sensitized BRAF mutant and BRAF/NRAS wildtype CMM cells to TH1579. AXL overexpression culminated in increased ROS levels in CMM cells. Moreover, silencing of a protein that has shown opposing effects on cell proliferation, CAV-1, decreased sensitivity to TH1579 in a BRAF inhibitor resistant cell line. AXL-MTH1 and CAV-1-MTH1 mRNA expressions were correlated as seen in CMM clinical samples. Finally, TH1579 in combination with BRAF inhibitor exhibited a more potent cell killing effect in BRAF mutant cells both in vitro and in vivo. In summary, we show that TH1579-mediated efficacy is independent of BRAF/NRAS mutational status but dependent on the expression of AXL and CAV-1.

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“…Recently, a compound initially designed as MTH1 (Mut T homolog 1) inhibitor, TH588, was shown to dock into the colchicine-binding pocket [176]. By reducing microtubule plus end dynamics, this cyclopropyl analog affects tubulin polymerization, resulting in disruption of mitotic spindles, prolongation of mitosis and, eventually, apoptosis [176][177][178]. Preclinical studies show promising results for the use of TH588 as an anticancer drug [179,180].…”

Section: Colchicine Sitementioning

confidence: 99%

Intrinsic and Extrinsic Factors Affecting Microtubule Dynamics in Normal and Cancer Cells

et al. 2020

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Microtubules (MTs), highly dynamic structures composed of α- and β-tubulin heterodimers, are involved in cell movement and intracellular traffic and are essential for cell division. Within the cell, MTs are not uniform as they can be composed of different tubulin isotypes that are post-translationally modified and interact with different microtubule-associated proteins (MAPs). These diverse intrinsic factors influence the dynamics of MTs. Extrinsic factors such as microtubule-targeting agents (MTAs) can also affect MT dynamics. MTAs can be divided into two main categories: microtubule-stabilizing agents (MSAs) and microtubule-destabilizing agents (MDAs). Thus, the MT skeleton is an important target for anticancer therapy. This review discusses factors that determine the microtubule dynamics in normal and cancer cells and describes microtubule–MTA interactions, highlighting the importance of tubulin isoform diversity and post-translational modifications in MTA responses and the consequences of such a phenomenon, including drug resistance development.

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VISAGE Reveals a Targetable Mitotic Spindle Vulnerability in Cancer Cells

Cited by 24 publications

References 92 publications

Pharmaceutical-grade Rigosertib is a Microtubule-destabilizing Agent

Pharmaceutical-grade Rigosertib is a Microtubule-destabilizing Agent

AXL and CAV-1 play a role for MTH1 inhibitor TH1579 sensitivity in cutaneous malignant melanoma

Intrinsic and Extrinsic Factors Affecting Microtubule Dynamics in Normal and Cancer Cells

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