Principal Investigator

Xiaolin
Zi
Awardee Organization

University Of California-Irvine
United States

Fiscal Year
2020
Activity Code
R01
Project End Date

The NEDD8 pathway mediated Skp2 degradation in chemoprevention by FKA

Chemoprevention in unselected populations has been largely unsuccessful; therefore targeting chemoprevention to high-risk populations by molecular subtyping may represent a promising future strategy. Flavokawain A (FKA) is a naturally occurring chalcone identified from the kava plant. Epidemiological studies have suggested that kava consumption may be associated with much lower prostate cancer (PCa) incidence in the three highest kava-drinking countries (Vanuatu, Fiji, and Western Samoa). We have shown that FKA selectively inhibits the growth of PCa cell lines and Rb deficient cells compared to normal cells. In addition, dietary feeding of FKA inhibited the formation of high-grade prostatic intra-epithelial neoplasia lesions and prostate adenocarcinomas, reduced the tumor burden and completely abolished distant organ metastasis.in the TRAMP transgenic mouse model without showing any toxicity. The mechanism of FKA’s action, in particular the anti-cancer molecular target (s) of FKA, remains largely unknown. We have therefore carried out extensive preliminary studies. We found that (1) FKA is five times more effective in inhibiting the growth of PC3 cells with over-expression of Skp2 (the IC50 is about 8 M) than PC3 cells expressing a vector control; (2) FKA accelerates Skp2 degradation via a proteasome and ubiquitination dependent pathway; (3) Skp2 degradation by FKA is not dependent on Cdh1 expression but associated with functional Cullin1; (4) FKA inhibits the conjugation of NEDD8 to Uba3 and Ubc12 in an in vitro assay and in cell culture; and (5) Given FKA has α, β-unsaturated carbonyl moiety , FKA may react with SH groups of cysteine in the Uba3 and Ubc12. A focused library of FKA analogs and α, β-unsaturated carbonyl compounds has therefore been prepared for a structure-activity relationship study. Based on these preliminary studies, we hypothesize that FKA functions as a novel inhibitor of NEDD8 conjugation and causes Skp2 ubiquitination and degradation; and that FKA exhibits chemopreventive activity by causing the accumulation of Skp2 substrates (i.e. p27, FOXO1, -TrCP, etc.) leading to cell cycle arrest and apoptosis. To test the hypotheses, we will determine: (i) The mechanisms by which FKA inhibits the process of NEDD8 conjugation; (ii) Whether inhibition of NEDD8 conjugation by FKA is causally associated with Skp2 degradation; (iii) The active concentrations of FKA or FKA metabolites in the mouse prostate and plasma that can be achieved for effective inhibition of NEDD8 conjugation and for down-regulation Skp2; (vi) Whether inhibition of the targets (i.e. NEDD8 and Skp2) by FKA at its effective concentrations will prevent or delay prostate carcinogenesis; (v) Whether more potent Skp2 targeting agents can be identified by studying the structure-activity relationship of FKA analogs and α, β-unsaturated carbonyl compounds in deNEDDylation and Skp2 degradation. Since pRb and Pten loss are common in PCa and Skp2 has been shown to be a major oncogenic target during the process of the pRb or Pten loss initiated cancer, these molecules present attractive targets for chemoprevention. Answers to the questions described above would bring novel impact on PCa prevention by defining a novel compound targeting the tumor suppressor pRb or Pten loss mediated PCa initiation pathway.

Publications

  • Xu X, Tian X, Song L, Xie J, Liao JC, Meeks JJ, Wu XR, Gin GE, Wang B, Uchio E, Zi X. Kawain Inhibits Urinary Bladder Carcinogenesis through Epigenetic Inhibition of LSD1 and Upregulation of H3K4 Methylation. Biomolecules. 2023 Mar 13;13. (3). PMID: 36979456
  • Liu Z, Song L, Xie J, Wu XR, Gin GE, Wang B, Uchio E, Zi X. Kavalactone Kawain Impedes Urothelial Tumorigenesis in UPII-Mutant Ha-Ras Mice via Inhibition of mTOR Signaling and Alteration of Cancer Metabolism. Molecules (Basel, Switzerland). 2023 Feb 9;28. (4). PMID: 36838656
  • Liu Z, Song L, Xie J, Simoneau AR, Uchio E, Zi X. Chemoprevention of Urothelial Cell Carcinoma Tumorigenesis by Dietary Flavokawain A in UPII-Mutant Ha-ras Transgenic Mice. Pharmaceutics. 2022 Feb 24;14. (3). PMID: 35335874
  • Li X, Song L, Xu S, Tippin M, Meng S, Xie J, Uchio E, Zi X. Kava root extracts hinder prostate cancer development and tumorigenesis by involvement of dual inhibition of MAO-A and LSD1. Journal of translational genetics and genomics. 2021;5:163-172. Epub 2021 May 28. PMID: 34368644
  • Li Y, Pham V, Bui M, Song L, Wu C, Walia A, Uchio E, Smith-Liu F, Zi X. Rhodiola rosea L.: an herb with anti-stress, anti-aging, and immunostimulating properties for cancer chemoprevention. Current pharmacology reports. 2017 Dec;3(6):384-395. Epub 2017 Sep 14. PMID: 30393593
  • Fu DJ, Zhao RH, Li JH, Yang JJ, Mao RW, Wu BW, Li P, Zi XL, Zhang QQ, Cai HJ, Zhang SY, Zhang YB, Liu HM. Molecular diversity of phenothiazines: design and synthesis of phenothiazine-dithiocarbamate hybrids as potential cell cycle blockers. Molecular diversity. 2017 Nov;21(4):933-942. Epub 2017 Aug 7. PMID: 28785928
  • Jandial DD, Krill LS, Chen L, Wu C, Ke Y, Xie J, Hoang BH, Zi X. Induction of G2M Arrest by Flavokawain A, a Kava Chalcone, Increases the Responsiveness of HER2-Overexpressing Breast Cancer Cells to Herceptin. Molecules (Basel, Switzerland). 2017 Mar 14;22. (3). PMID: 28335434
  • Zhang S, Li T, Zhang L, Wang X, Dong H, Li L, Fu D, Li Y, Zi X, Liu HM, Zhang Y, Xu H, Jin CY. A novel chalcone derivative S17 induces apoptosis through ROS dependent DR5 up-regulation in gastric cancer cells. Scientific reports. 2017 Aug 29;7(1):9873. PMID: 28852176
  • Liu Z, Ha US, Yu K, Wu C, Yokoyama N, Zi X. Kavalactone yangonin induces autophagy and sensitizes bladder cancer cells to flavokawain A and docetaxel via inhibition of the mTOR pathway. Journal of biomedical research. 2017 Sep 26;31(5):408-418. PMID: 28959001
  • Chen LX, Xia GY, He H, Huang J, Qiu F, Zi XL. New withanolides with TRAIL-sensitizing effect from Physalis pubescens L. RSC advances. 2016;6(58):52925-52936. Epub 2016 May 26. PMID: 27822364
  • Wang J, Aldahamsheh O, Ferrena A, Borjihan H, Singla A, Yaguare S, Singh S, Viscarret V, Tingling J, Zi X, Lo Y, Gorlick R, Zheng D, Schwartz EL, Zhao H, Yang R, Geller DS, Hoang BH. The interaction of SKP2 with p27 enhances the progression and stemness of osteosarcoma. Annals of the New York Academy of Sciences. 2021 Apr;1490(1):90-104. Epub 2021 Feb 16. PMID: 33594717
  • Zhang S, Li T, Zhang Y, Xu H, Li Y, Zi X, Yu H, Li J, Jin CY, Liu HM. A new brominated chalcone derivative suppresses the growth of gastric cancer cells in vitro and in vivo involving ROS mediated up-regulation of DR5 and 4 expression and apoptosis. Toxicology and applied pharmacology. 2016 Oct 15;309:77-86. Epub 2016 Sep 2. PMID: 27594528
  • Li X, Pham V, Tippin M, Fu D, Rendon R, Song L, Uchio E, Hoang BH, Zi X. Flavokawain B targets protein neddylation for enhancing the anti-prostate cancer effect of Bortezomib via Skp2 degradation. Cell communication and signaling : CCS. 2019 Mar 18;17(1):25. PMID: 30885218
  • Zhang Y, Zvi YS, Batko B, Zaphiros N, O'Donnell EF, Wang J, Sato K, Yang R, Geller DS, Koirala P, Zhang W, Du X, Piperdi S, Liu Y, Zheng D, Roth M, Gill J, Zhang J, Ren T, Gorlick R, Zi X, Hoang BH. Down-regulation of Skp2 expression inhibits invasion and lung metastasis in osteosarcoma. Scientific reports. 2018 Sep 24;8(1):14294. PMID: 30250282
  • Pham V, Rendon R, Le VX, Tippin M, Fu DJ, Le TH, Miller M, Agredano E, Cedano J, Zi X. Gartanin is a novel NEDDylation inhibitor for induction of Skp2 degradation, FBXW2 expression, and autophagy. Molecular carcinogenesis. 2020 Feb;59(2):193-201. Epub 2019 Nov 29. PMID: 31782573
  • Song L, Mino M, Yamak J, Nguyen V, Lopez D, Pham V, Fazelpour A, Le V, Fu D, Tippin M, Uchio E, Zi X. Flavokawain A Reduces Tumor-Initiating Properties and Stemness of Prostate Cancer. Frontiers in oncology. 2022 Jul 13;12:943846. doi: 10.3389/fonc.2022.943846. eCollection 2022. PMID: 35912174
  • Khajeh NR, Khoyilar C, Wu Y, Spradling K, Zi X, Youssef RF. Bladder Cancer Chemopreventive Agents: Current Knowledge and Concepts. Mini reviews in medicinal chemistry. 2018;18(13):1143-1150. PMID: 26975668
  • Ma YC, Ke Y, Zi X, Zhao F, Yuan L, Zhu YL, Fan XX, Zhao NM, Li QY, Qin YH, Liu HM. Induction of the mitochondria-mediated apoptosis in human esophageal cancer cells by DS2, a newly synthetic diterpenoid analog, is regulated by Bax and caused by generation of reactive oxygen species. Oncotarget. 2016 Dec 27;7(52):86211-86224. PMID: 27863415
  • Fu DJ, Zhang L, Song J, Mao RW, Zhao RH, Liu YC, Hou YH, Li JH, Yang JJ, Jin CY, Li P, Zi XL, Liu HM, Zhang SY, Zhang YB. Design and synthesis of formononetin-dithiocarbamate hybrids that inhibit growth and migration of PC-3 cells via MAPK/Wnt signaling pathways. European journal of medicinal chemistry. 2017 Feb 15;127:87-99. Epub 2016 Dec 14. PMID: 28038329
  • Fu DJ, Song J, Hou YH, Zhao RH, Li JH, Mao RW, Yang JJ, Li P, Zi XL, Li ZH, Zhang QQ, Wang FY, Zhang SY, Zhang YB, Liu HM. Discovery of 5,6-diaryl-1,2,4-triazines hybrids as potential apoptosis inducers. European journal of medicinal chemistry. 2017 Sep 29;138:1076-1088. Epub 2017 Jul 8. PMID: 28763643
  • Chen L, Xia G, Qiu F, Wu C, Denmon AP, Zi X. Physapubescin selectively induces apoptosis in VHL-null renal cell carcinoma cells through down-regulation of HIF-2α and inhibits tumor growth. Scientific reports. 2016 Sep 1;6:32582. PMID: 27581364