Targeting E2 ubiquitin-conjugating enzyme UbcH5c by small molecule inhibitor suppresses pancreatic cancer growth and metastasis

Molecular Cancer - Tập 21 Số 1
Simin Qi1, Xiaoqing Guan1, Jia Zhang2, Dehua Yu1, Xiaoduo Yu1, Qinglin Li1, Wenjuan Yin1, Xiangdong Cheng1, Weidong Zhang3, Jiang‐Jiang Qin4
1The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou, 310022, Zhejiang, China
2Shanxi Institute of Traditional Chinese Medicine, Taiyuan, 030012, China
3Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
4College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, 310053, China

Tóm tắt

Abstract Background Pancreatic cancer is one of the most lethal cancers worldwide. The IAPs function as E3 ubiquitin ligases and contribute to pancreatic cancer initiation, progression, and metastasis. Although IAP-targeted therapies have been developed and shown anticancer efficacy in preclinical settings, none of them has been approved yet. Methods Transcriptome data from public datasets were used to analyze the correlation of IAPs and E2s, and the biological function of E2 UbcH5c in pancreatic cancer. A structure-based virtual screen was used to identify UbcH5c inhibitor, and surface plasmon resonance analysis and cellular thermal shift assays were employed to evaluate the binding affinity. The anticancer activities were demonstrated through in vitro and in vivo assays, while the related mechanisms were explored through transcriptomic and proteomic analyses and confirmed by western blot, immunofluorescence, and qRT-PCR. Results UbcH5c is positively correlated with the expression of IAPs in pancreatic cancer. We further found that UbcH5c is overexpressed and associated with a poor prognosis in pancreatic cancer. We identified a small-molecule UbcH5c inhibitor, termed DHPO, which directly bound to UbcH5c protein. DHPO inhibited cell viability and colony formation, induced apoptosis, and suppressed migration and invasion of pancreatic cancer cells in vitro. The compound inhibited UbcH5c-mediated IκBα degradation and NF-κB activation, which is critical for its anticancer activity. Furthermore, DHPO suppressed the tumor growth and metastasis in two orthotopic pancreatic tumor mouse models. Conclusions These results indicated that inhibiting UbcH5c is a novel and effective strategy for treating pancreatic cancer and DHPO represents a new class of UbcH5c inhibitor and may be further developed as an anti-pancreatic cancer therapeutic agent.

Từ khóa


Tài liệu tham khảo

Mizrahi JD, Surana R, Valle JW, Shroff RT. Pancreatic cancer. Lancet. 2020;395:2008–20.

Strobel O, Neoptolemos J, Jäger D, Büchler MW. Optimizing the outcomes of pancreatic cancer surgery. Nat Rev Clin Oncol. 2019;16:11–26.

Neoptolemos JP, Kleeff J, Michl P, Costello E, Greenhalf W, Palmer DH. Therapeutic developments in pancreatic cancer: current and future perspectives. Nat Rev Gastroenterol Hepatol. 2018;15:333–48.

Christenson ES, Jaffee E, Azad NS. Current and emerging therapies for patients with advanced pancreatic ductal adenocarcinoma: a bright future. Lancet Oncol. 2020;21:e135–45.

Golan T, Hammel P, Reni M, Van Cutsem E, Macarulla T, Hall MJ, et al. Maintenance olaparib for germline BRCA-mutated metastatic pancreatic cancer. N Engl J Med. 2019;381:317–27.

Deng L, Meng T, Chen L, Wei W, Wang P. The role of ubiquitination in tumorigenesis and targeted drug discovery. Signal Transduct Target Ther. 2020;5:11.

Celebi G, Kesim H, Ozer E, Kutlu O. The effect of dysfunctional ubiquitin enzymes in the pathogenesis of most common diseases. Int J Mol Sci. 2020;21:6335.

Berndsen CE, Wolberger C. New insights into ubiquitin E3 ligase mechanism. Nat Struct Mol Biol. 2014;21:301–7.

Cong H, Xu L, Wu Y, Qu Z, Bian T, Zhang W, et al. Inhibitor of apoptosis protein (IAP) antagonists in anticancer agent discovery: Current status and perspectives. J Med Chem. 2019;62:5750–72.

Qi SM, Cheng G, Cheng XD, Xu Z, Xu B, Zhang WD, et al. Targeting USP7-mediated deubiquitination of MDM2/MDMX-p53 pathway for cancer therapy: Are we there yet? Front Cell Dev Biol. 2020;8:233.

Wang W, Qin JJ, Rajaei M, Li X, Yu X, Hunt C, et al. Targeting MDM2 for novel molecular therapy: Beyond oncology. Med Res Rev. 2020;40:856–80.

Kumar S, Fairmichael C, Longley DB, Turkington RC. The multiple roles of the IAP super-family in cancer. Pharmacol Ther. 2020;214:107610.

Gao X, Zhang L, Wei Y, Yang Y, Li J, Wu H, et al. Prognostic value of XIAP level in patients with various cancers: A systematic review and meta-analysis. J Cancer. 2019;10:1528–37.

Li S, Sun J, Yang J, Zhang L, Wang L, Wang X, et al. XIAP expression is associated with pancreatic carcinoma outcome. Mol Clin Oncol. 2013;1:305–8.

Blaquiere N, Villemure E, Staben ST. Medicinal chemistry of inhibiting RING-type E3 ubiquitin ligases. J Med Chem. 2020;63:7957–85.

Du X, Song H, Shen N, Hua R, Yang G. The molecular basis of ubiquitin-conjugating enzymes (E2s) as a potential target for cancer therapy. Int J Mol Sci. 2021;22:3440.

Zhao ZK, Wu WG, Chen L, Dong P, Gu J, Mu JS, et al. Expression of UbcH10 in pancreatic ductal adenocarcinoma and its correlation with prognosis. Tumour Biol. 2013;34:1473–7.

Wang X, Yin L, Yang L, Zheng Y, Liu S, Yang J, et al. Silencing ubiquitin-conjugating enzyme 2C inhibits proliferation and epithelial-mesenchymal transition in pancreatic ductal adenocarcinoma. FEBS J. 2019;286:4889–909.

Zheng YW, Gao PF, Ma MZ, Chen Y, Li CY. Role of ubiquitin-conjugating enzyme E2T in the carcinogenesis and progression of pancreatic cancer. Oncol Lett. 2020;20:1462–8.

Wang L, Liang Y, Li P, Liang Q, Sun H, Xu D, et al. Oncogenic activities of UBE2S mediated by VHL/HIF-1alpha/STAT3 signal via the ubiquitin-proteasome system in PDAC. Onco Targets Ther. 2019;12:9767–81.

Shang M, Weng L, Xu G, Wu S, Liu B, Yin X, et al. TRIM11 suppresses ferritinophagy and gemcitabine sensitivity through UBE2N/TAX1BP1 signaling in pancreatic ductal adenocarcinoma. J Cell Physiol. 2021;236:6868–83.

Nie LY, Qin JJ, Huang Y, Yan L, Liu YB, Pan YX, et al. Sesquiterpenoids from Inula lineariifolia inhibit nitric oxide production. J Nat Prod. 2010;73:1117–20.

Wang W, Qin JJ, Voruganti S, Nijampatnam B, Velu SE, Ruan KH, et al. Discovery and characterization of dual inhibitors of MDM2 and NFAT1 for pancreatic cancer therapy. Cancer Res. 2018;78:5656–67.

Wang W, Qin JJ, Voruganti S, Wang MH, Sharma H, Patil S, et al. Identification of a new class of MDM2 inhibitor that inhibits growth of orthotopic pancreatic tumors in mice. Gastroenterology. 2014;147:893–902.

Wang W, Qin JJ, Voruganti S, Srivenugopal KS, Nag S, Patil S, et al. The pyrido[b]indole MDM2 inhibitor SP-141 exerts potent therapeutic effects in breast cancer models. Nat Commun. 2014;5:5086.

Wang W, Cheng JW, Qin JJ, Hu B, Li X, Nijampatnam B, et al. MDM2-NFAT1 dual inhibitor, MA242: Effective against hepatocellular carcinoma, independent of p53. Cancer Lett. 2019;459:156–67.

Zhang J, Wang W, Zhou Y, Yang J, Xu J, Xu Z, et al. Terphenyllin suppresses orthotopic pancreatic tumor growth and prevents metastasis in mice. Front Pharmacol. 2020;11:457.

Guan X, Cai M, Du Y, Yang E, Ji J, Wu J. CVCDAP: an integrated platform for molecular and clinical analysis of cancer virtual cohorts. Nucleic Acids Res. 2020;48:W463–71.

Zhou Y, Chen R, Luo X, Zhang WD, Qin JJ. The E2 ubiquitin-conjugating enzyme UbcH5c: an emerging target in cancer and immune disorders. Drug Discov Today. 2020;25:1988–97.

Liu L, Hua Y, Wang D, Shan L, Zhang Y, Zhu J, et al. A sesquiterpene lactone from a medicinal herb inhibits proinflammatory activity of TNF-alpha by inhibiting ubiquitin-conjugating enzyme UbcH5. Chem Biol. 2014;21:1341–50.

Qin JJ, Wang W, Voruganti S, Wang H, Zhang WD, Zhang R. Identification of a new class of natural product MDM2 inhibitor: In vitro and in vivo anti-breast cancer activities and target validation. Oncotarget. 2015;6:2623–40.

Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005;102:15545–50.

Zhou Y, Zhou B, Pache L, Chang M, Khodabakhshi AH, Tanaseichuk O, et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nat Commun. 2019;10:1523.

Ho WJ, Jaffee EM, Zheng L. The tumour microenvironment in pancreatic cancer - clinical challenges and opportunities. Nat Rev Clin Oncol. 2020;17:527–40.

Hosein AN, Brekken RA, Maitra A. Pancreatic cancer stroma: an update on therapeutic targeting strategies. Nat Rev Gastroenterol Hepatol. 2020;17:487–505.

Collisson EA, Bailey P, Chang DK, Biankin AV. Molecular subtypes of pancreatic cancer. Nat Rev Gastroenterol Hepatol. 2019;16:207–20.

Zhang S, Zhou Y, Sarkeshik A, Yates JR, Thomson TM, Zhang Z, et al. Identification of RNF8 as a ubiquitin ligase involved in targeting the p12 subunit of DNA polymerase δ for degradation in response to DNA damage. J Biol Chem. 2013;288:2941–50.

Nara M, Teshima K, Watanabe A, Ito M, Iwamoto K, Kitabayashi A, et al. Bortezomib reduces the tumorigenicity of multiple myeloma via downregulation of upregulated targets in clonogenic side population cells. PLoS One. 2013;8:e56954.

Huang XH, Yan X, Zhang QH, Hong P, Zhang WX, Liu YP, et al. Direct targeting of HSP90 with daurisoline destabilizes β-catenin to suppress lung cancer tumorigenesis. Cancer Lett. 2020;489:66–78.

Chen H, Wu G, Gao S, Guo R, Zhao Z, Yuan H, et al. Discovery of potent small-molecule inhibitors of ubiquitin-conjugating enzyme UbcH5c from α-santonin derivatives. J Med Chem. 2017;60:6828–52.

Huang M, Zhou Y, Duan D, Yang C, Zhou Z, Li F, et al. Targeting ubiquitin conjugating enzyme UbcH5b by a triterpenoid PC3-15 from Schisandra plants sensitizes triple-negative breast cancer cells to lapatinib. Cancer Lett. 2021;504:125–36.

Qin JJ, Jin HZ, Huang Y, Zhang SD, Shan L, Voruganti S, et al. Selective cytotoxicity, inhibition of cell cycle progression, and induction of apoptosis in human breast cancer cells by sesquiterpenoids from Inula lineariifolia Turcz. Eur J Med Chem. 2013;68:473–81.

Wu G, Zhu L, Yuan X, Chen H, Xiong R, Zhang S, et al. Britanin ameliorates cerebral ischemia-reperfusion injury by inducing the Nrf2 protective pathway. Antioxid Redox Signal. 2017;27:754–68.

Taniguchi K, Karin M. NF-κB, inflammation, immunity and cancer: coming of age. Nat Rev Immunol. 2018;18:309–24.

Tundo GR, Sbardella D, Santoro AM, Coletta A, Oddone F, Grasso G, et al. The proteasome as a druggable target with multiple therapeutic potentialities: Cutting and non-cutting edges. Pharmacol Ther. 2020;213:107579.

Antonia RJ, Hagan RS, Baldwin AS. Expanding the view of IKK: New substrates and new biology. Trends Cell Biol. 2021;31:166–78.

Eluard B, Thieblemont C, Baud V. NF-κB in the new era of cancer therapy. Trends Cancer. 2020;6:677–87.

Li Y, Sun XX, Elferich J, Shinde U, David LL, Dai MS. Monoubiquitination is critical for ovarian tumor domain-containing ubiquitin aldehyde binding protein 1 (Otub1) to suppress UbcH5 enzyme and stabilize p53 protein. J Biol Chem. 2014;289:5097–108.

Peng H, Yang J, Li G, You Q, Han W, Li T, et al. Ubiquitylation of p62/sequestosome1 activates its autophagy receptor function and controls selective autophagy upon ubiquitin stress. Cell Res. 2017;27:657–74.

Yang N, Chen T, Wang L, Liu R, Niu Y, Sun L, et al. CXCR4 mediates matrix stiffness-induced downregulation of UBTD1 driving hepatocellular carcinoma progression via YAP signaling pathway. Theranostics. 2020;10:5790–801.

Thông tin
Thông tin xuất bản

Molecular Cancer

Tập 21 Số 1

Thông tin tác giả