(no more than 30 lines) Aberrant ubiquitination processes are tightly associated with various human diseases including cancer. The Anaphase Promoting Complex (APC) has been documented to regulate cell cycle progression through binding activator proteins Cdh1 and Cdc20 at different cell cycle stages to promote APC-dependent ubiquitination and destruction of their substrates. Recent studies revealed that APCCdc20 is an attractive anti-cancer target and depletion of Cdc20 suppresses in vivo tumorigenesis in part by triggering mitotic arrest and subsequent apoptosis. We have recently reported the pro-apoptotic protein Bim as a novel APCCdc20 substrate, which supports the notion that Cdc20 is an oncogene. However, how Cdc20 is controlled by upstream regulator(s) and how the Cdc20-Bim signaling axis contributes to chemoradio-resistance in vivo remains largely unaddressed. Our preliminary results showed that the CUL3SPOP E3 ligase specifically targets Cdc20 for ubiquitination and subsequent destruction to attenuate its oncogenic activity. Furthermore, we demonstrated that APCCdc20 governs the ubiquitination and subsequent destruction of Bim in a D-box-dependent manner. Notably, human Adult-T-cell-Leukemia cells that acquire elevated APCCdc20 activity via expressing the Tax-viral- oncoprotein, exhibit reduced Bim levels and corresponding resistance to anti-cancer reagents. Conversely, depletion of Cdc20 in various human cancer cell lines sensitized their response to various chemotherapeutic drugs. More importantly, Cdc20 and multiple APC-core components were identified in an siRNA screen that upon knockdown sensitizes head-and-neck-cancer cells to ?-irradiation in a Bim-dependent manner. Based on our preliminary data, we hypothesize that SPOP is a novel upstream negative regulator of Cdc20 stability and oncogenic functions, and APCCdc20 negatively governs the induction of apoptotic pathway in response to anti- mitotic agents and ?-irradiation in part through targeting Bim for ubiquitination and destruction. We will examine our hypothesis by accomplishing the following three specific aims.
In Aim #1, we will determine the molecular mechanisms underlying SPOP-mediated ubiqutination and degradation of Cdc20 and to examine whether SPOP deficiency in head and neck cancer leads to elevated tumorigenesis in part by Cdc20 accumulation. Our proposed work in Aim #2 will determine the physiological role of APCCdc20 in governing cellular apoptosis in vivo by regulating the abundance of Bim and other Cdc20 substrates. These proposed studies would help to further understand the molecular mechanisms underlying anti-mitotic agents-induced apoptosis, and provide the rationale for developing specific Cdc20 inhibitors as effective anti-cancer therapies.
In Aim #3, we will validate the APCCdc20 E3 ligase as a novel radio-sensitizing target. Taken together, these studies will significantly extend our current knowledge of how depletion of Cdc20 exerts its anti-tumor biological functions through controlling the abundance of Bim. Our proposed studies will provide the molecular basis for designing novel strategies to target Cdc20 by radiation for achieving better clinical treatment of cancer patients.
This project is focused on characterizing SPOP as a novel upstream negative regulator of Cdc20 stability, as well as revealing the physiological role of APCCdc20 in cellular apoptosis regulation by targeting the pro-apoptotic protein, Bim, for ubiquitination and destruction. We will use both in vitro molecular biology approaches and in vivo animal models to characterize the molecular mechanisms by which Cdc20 functions as an oncogene through promoting the destruction of the Bim tumor suppressor. By using both genetic and pharmacological approaches under physiological relevant in vivo settings, the proposed studies will validate mechanistically that APCCdc20 is an attractive anti- cancer target, thus offering the molecular basis and rationale for future development of APC inhibitors (such as pro-TAME or Apcin) as a novel class of chemoradiation sensitizing agents.
|Zhang, Jinfang; Bu, Xia; Wang, Haizhen et al. (2018) Cyclin D-CDK4 kinase destabilizes PD-L1 via cullin 3-SPOP to control cancer immune surveillance. Nature 553:91-95|
|Tzoneva, Gannie; Dieck, Chelsea L; Oshima, Koichi et al. (2018) Clonal evolution mechanisms in NT5C2 mutant-relapsed acute lymphoblastic leukaemia. Nature 553:511-514|
|Wan, Lixin; Chen, Ming; Cao, Juxiang et al. (2017) The APC/C E3 Ligase Complex Activator FZR1 Restricts BRAF Oncogenic Function. Cancer Discov 7:424-441|
|Wu, Fei; Dai, Xiangpeng; Gan, Wenjian et al. (2017) Prostate cancer-associated mutation in SPOP impairs its ability to target Cdc20 for poly-ubiquitination and degradation. Cancer Lett 385:207-214|
|Dai, Xiangpeng; Gan, Wenjian; Li, Xiaoning et al. (2017) Prostate cancer-associated SPOP mutations confer resistance to BET inhibitors through stabilization of BRD4. Nat Med 23:1063-1071|
|Wang, Bin; Jie, Zuliang; Joo, Donghyun et al. (2017) TRAF2 and OTUD7B govern a ubiquitin-dependent switch that regulates mTORC2 signalling. Nature 545:365-369|
|Shimizu, Kouhei; Fukushima, Hidefumi; Ogura, Kohei et al. (2017) The SCF?-TRCP E3 ubiquitin ligase complex targets Lipin1 for ubiquitination and degradation to promote hepatic lipogenesis. Sci Signal 10:|
|Fukushima, Hidefumi; Shimizu, Kouhei; Watahiki, Asami et al. (2017) NOTCH2 Hajdu-Cheney Mutations Escape SCFFBW7-Dependent Proteolysis to Promote Osteoporosis. Mol Cell 68:645-658.e5|
|Nihira, Naoe T; Ogura, Kohei; Shimizu, Kouhei et al. (2017) Acetylation-dependent regulation of MDM2 E3 ligase activity dictates its oncogenic function. Sci Signal 10:|
|Cheng, Ji; Zhang, Tao; Ji, Hongbin et al. (2016) Functional characterization of AMP-activated protein kinase signaling in tumorigenesis. Biochim Biophys Acta 1866:232-251|
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