Small-molecule inhibition of Wee1 kinase by MK-1775 selectively sensitizes p53-deficient tumor cells to DNA-damaging agents

Hirai, Hiroshi; Iwasawa, Yoshikazu; Okada, Megumu; Arai, T.; Nishibata, Toshihide; Kobayashi, Makiko; Kimura, Toshifumi; Kaneko, Naoki; Ohtani, Junko; Yamanaka, Kazunori; Itadani, Hiraku; Takahashi-Suzuki, Ikuko; Fukasawa, Kazuhiro; Oki, Hiroko; Nambu, Tadahiro; Jiang, Jian; Sakai, Takumi; Arakawa, Hiroharu; Sakamoto, Toshihiro; Sagara, Takeshi; Yoshizumi, Takashi; Mizuarai, Shinji; Kotani, Hidehito

doi:10.1158/1535-7163.mct-09-0463

Cited by 460 publications

(530 citation statements)

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“…The consensus view is that Wee1 inhibition facilitates tumor cell killing through G 2 /M checkpoint inactivation, which would catalyze mitotic catastrophe (23)(24)(25). As expected, Wee1 inhibition is synergistic with DNA-damaging agents, specifically in TP53-mutant cancer cell lines (3). However, a recent study indicated that only a subset of TP53-mutant patient-derived pancreatic cancer xenografts showed benefit from Wee1 inhibition (21).…”

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confidence: 77%

“…Chemical inhibitors for several cell cycle checkpoint kinases have been developed. Preclinical research, however, has shown that the efficacy of therapeutic checkpoint inhibition is context-sensitive and depends on the genetic make-up of an individual cancer (2,3). Therefore, to optimally implement such novel inhibitors in the clinic, the molecular characteristics that determine inhibitor activity need to be identified to select eligible patients and to anticipate on mechanisms of acquired resistance.…”

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confidence: 99%

“…Wee1 is one of the G 2 /M cell cycle checkpoint kinases for which selective chemical inhibitors have been developed as anticancer drugs (3,18). Inhibition of Wee1 potentiates DNA-damaging chemotherapeutics and radiotherapy and has cytotoxic effects as a single agent (19)(20)(21)(22).…”

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confidence: 99%

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A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

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Blomen

Bisteau

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The Wee1 cell cycle checkpoint kinase prevents premature mitotic entry by inhibiting cyclin-dependent kinases. Chemical inhibitors of Wee1 are currently being tested clinically as targeted anticancer drugs. Wee1 inhibition is thought to be preferentially cytotoxic in p53-defective cancer cells. However, TP53 mutant cancers do not respond consistently to Wee1 inhibitor treatment, indicating the existence of genetic determinants of Wee1 inhibitor sensitivity other than TP53 status. To optimally facilitate patient selection for Wee1 inhibition and uncover potential resistance mechanisms, identification of these currently unknown genes is necessary. The aim of this study was therefore to identify gene mutations that determine Wee1 inhibitor sensitivity. We performed a genome-wide unbiased functional genetic screen in TP53 mutant near-haploid KBM-7 cells using gene-trap insertional mutagenesis. Insertion site mapping of cells that survived long-term Wee1 inhibition revealed enrichment of G 1 /S regulatory genes, including SKP2, CUL1, and CDK2. Stable depletion of SKP2, CUL1, or CDK2 or chemical Cdk2 inhibition rescued the γ-H2AX induction and abrogation of G 2 phase as induced by Wee1 inhibition in breast and ovarian cancer cell lines. Remarkably, live cell imaging showed that depletion of SKP2, CUL1, or CDK2 did not rescue the Wee1 inhibition-induced karyokinesis and cytokinesis defects. These data indicate that the activity of the DNA replication machinery, beyond TP53 mutation status, determines Wee1 inhibitor sensitivity, and could serve as a selection criterion for Wee1-inhibitor eligible patients. Conversely, loss of the identified S-phase genes could serve as a mechanism of acquired resistance, which goes along with development of severe genomic instability.cell cycle | checkpoint | AZD-1775 | MK-1775 | polyploidy

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A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

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Blomen

Bisteau

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…18 Subsequently it was reported that treatment with the clinical Wee1 inhibitor MK-1775 (AZD-1775) likewise had striking cytokinetic effects, including features suggestive of premature entry into mitosis during early S-phase. 19,20 Of the Wee1 inhibitors, MK-1775 is currently the most advanced in clinical development, and it is reported to be highly selective toward Wee1 (21). As a single agent it appears to be well tolerated, but has little anti-cancer activity.…”

Section: Introductionmentioning

confidence: 99%

Cytokinetic effects of Wee1 disruption in pancreatic cancer

et al. 2016

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The Wee1 kinase, which is activated in response to DNA damage, regulates exit from G2 through inhibitory phosphorylation of Cdk1/Cdc2, and is an attractive drug target. However, recent work has highlighted effects of Cdk2 phosphorylation by Wee1 on movement through S-phase, suggesting the potential to sensitize to S-phase specific agents by Wee1 inhibitors. In this paper we applied multiparametric flow cytometry to patient-derived pancreatic cancer xenograft tumor cells to study the cell cycle perturbations of Wee1 disruption via the small molecule inhibitor MK-1775, and genetic knockdown. We find that in vitro treatment with MK-1775, and to a lesser degree, Wee1 RNA transcript knockdown, results in the striking appearance of S-phase cells prematurely entering into mitosis. This effect was not seen in vivo in any of the models tested. Here, although we noted an increase of S-phase cells expressing the damage response marker gH2AX, treatment with MK-1775 did not significantly sensitize cells to the cytidine analog gemcitabine. Treatment with MK-1775 did result in a transient but large increase in cells expressing the mitotic marker phosphorylated H3S10 that reached a peak 4 hours after treatment. This suggests a role for Wee1 regulating the progression of genomically unstable cancer cells through G2 in the absence of extrinsically-applied DNA damage. A single dose of 8Gy ionizing radiation resulted in the time-dependent accumulation of Cyclin A2 positive/phosphorylated H3S10 negative cells at the 4N position, which was abrogated by treatment with MK-1775. Consistent with these findings, a genomescale pooled RNA interference screen revealed that toxic doses of MK-1775 are suppressed by CDK2 or Cyclin A2 knockdown. These findings support G2 exit as the more significant effect of Wee1 inhibition in pancreatic cancers.

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“…Because p53 activation potently enforces the G 1 /S checkpoint, it has been theorized that p53-null tumors are especially dependent on the S and G 2 /M checkpoints to maintain genomic integrity. Therefore, inhibitors of cell cycle checkpoint proteins have largely been developed for the treatment of p53-deficient malignancies (5,21,24,47,48). Meanwhile, MDM2 i s have been developed for use in p53-sufficient malignancies because they have little or no impact in the absence of p53.…”

Section: Discussionmentioning

confidence: 99%

Manipulating DNA damage-response signaling for the treatment of immune-mediated diseases

McNally

Millen

Chaturvedi

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Antigen-activated lymphocytes undergo extraordinarily rapid cell division in the course of immune responses. We hypothesized that this unique aspect of lymphocyte biology leads to unusual genomic stress in recently antigen-activated lymphocytes and that targeted manipulation of DNA damage-response (DDR) signaling pathways would allow for selective therapeutic targeting of pathological T cells in disease contexts. Consistent with these hypotheses, we found that activated mouse and human T cells display a pronounced DDR in vitro and in vivo. Upon screening a variety of small-molecule compounds, we found that potentiation of p53 (via inhibition of MDM2) or impairment of cell cycle checkpoints (via inhibition of CHK1/2 or WEE1) led to the selective elimination of activated, pathological T cells in vivo. The combination of these strategies [which we termed "p53 potentiation with checkpoint abrogation" (PPCA)] displayed therapeutic benefits in preclinical disease models of hemophagocytic lymphohistiocytosis and multiple sclerosis, which are driven by foreign antigens or self-antigens, respectively. PPCA therapy targeted pathological T cells but did not compromise naive, regulatory, or quiescent memory T-cell pools, and had a modest nonimmune toxicity profile. Thus, PPCA is a therapeutic modality for selective, antigen-specific immune modulation with significant translational potential for diverse immune-mediated diseases.autoimmunity | immune regulation | DNA damage response | therapeutics T he immune system has evolved under intense pressure from pathogens that proliferate very rapidly (1). In responding to infections, the adaptive immune system needs to mobilize rare pathogen-specific lymphocytes quickly. This mobilization is achieved by rapid, exponential expansion of responding lymphocyte clones. Indeed, antigen-activated T and B cells appear to have some of the most rapid division rates among all mammalian cell types (2). We hypothesized that this unique aspect of lymphocyte biology would cause significant genomic stress in responding lymphocytes and that novel forms of therapeutic immune suppression could be developed by exploiting this weakness. Such strategies would allow for "developmental" or "kinetic" targeting of pathological immune responses at the time of disease activity or organ rejection and could complement or replace chronic suppression of immune signaling or cytokine pathways. The recent preclinical and clinical development of a wide array of rationally designed small-molecule inhibitors of various signaling and effector molecules within DNA damage-response (DDR) cascades has provided an opportunity to test this hypothesis (3, 4).We first looked for evidence of a DDR occurring in activated T cells in physiological contexts and found a broad DDR in murine and human T cells. Next, upon screening an array of DNA-damaging agents and DDR-altering small molecules for their ability to kill activated, but not resting, T cells, we identified two promising strategies. We found that inhibition of MDM2 (an end...

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Small-molecule inhibition of Wee1 kinase by MK-1775 selectively sensitizes p53-deficient tumor cells to DNA-damaging agents

Cited by 460 publications

References 28 publications

A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

Cytokinetic effects of Wee1 disruption in pancreatic cancer

Manipulating DNA damage-response signaling for the treatment of immune-mediated diseases

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Small-molecule inhibition of Wee1 kinase by MK-1775 selectively sensitizes p53-deficient tumor cells to DNA-damaging agents

Cited by 460 publications

References 28 publications

A haploid genetic screen identifies the G1/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

A haploid genetic screen identifies the G1/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

Cytokinetic effects of Wee1 disruption in pancreatic cancer

Manipulating DNA damage-response signaling for the treatment of immune-mediated diseases

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A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity

A haploid genetic screen identifies the G₁/S regulatory machinery as a determinant of Wee1 inhibitor sensitivity