{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T13:19:00Z","timestamp":1791638340844,"version":"4.3.4"},"reference-count":38,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,14]],"date-time":"2023-01-14T00:00:00Z","timestamp":1673654400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["T32GM008570"],"award-info":[{"award-number":["T32GM008570"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["R01GM134231"],"award-info":[{"award-number":["R01GM134231"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["R35GM128855"],"award-info":[{"award-number":["R35GM128855"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"NIH","doi-asserted-by":"publisher","award":["A17-026"],"award-info":[{"award-number":["A17-026"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Gabrielle\u2019s Angel Foundation Medical Research Award","award":["T32GM008570"],"award-info":[{"award-number":["T32GM008570"]}]},{"name":"Gabrielle\u2019s Angel Foundation Medical Research Award","award":["R01GM134231"],"award-info":[{"award-number":["R01GM134231"]}]},{"name":"Gabrielle\u2019s Angel Foundation Medical Research Award","award":["R35GM128855"],"award-info":[{"award-number":["R35GM128855"]}]},{"name":"Gabrielle\u2019s Angel Foundation Medical Research Award","award":["A17-026"],"award-info":[{"award-number":["A17-026"]}]},{"name":"University of North Carolina at Chapel Hill University Cancer Research Fund","award":["T32GM008570"],"award-info":[{"award-number":["T32GM008570"]}]},{"name":"University of North Carolina at Chapel Hill University Cancer Research Fund","award":["R01GM134231"],"award-info":[{"award-number":["R01GM134231"]}]},{"name":"University of North Carolina at Chapel Hill University Cancer Research Fund","award":["R35GM128855"],"award-info":[{"award-number":["R35GM128855"]}]},{"name":"University of North Carolina at Chapel Hill University Cancer Research Fund","award":["A17-026"],"award-info":[{"award-number":["A17-026"]}]},{"name":"Atomwise AIMS Award","award":["T32GM008570"],"award-info":[{"award-number":["T32GM008570"]}]},{"name":"Atomwise AIMS Award","award":["R01GM134231"],"award-info":[{"award-number":["R01GM134231"]}]},{"name":"Atomwise AIMS Award","award":["R35GM128855"],"award-info":[{"award-number":["R35GM128855"]}]},{"name":"Atomwise AIMS Award","award":["A17-026"],"award-info":[{"award-number":["A17-026"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Protein deubiquitinases play critical pathophysiological roles in cancer. Among all deubiquitinases, an oncogenic function for OTUD7B has been established in genetic NSCLC murine models. However, few deubiquitinase inhibitors have been developed due to technical challenges. Here, we report a putative small molecule OTUD7B inhibitor obtained from an AI-aided screen of a 4 million compound library. We validated the effects of the OTUD7B inhibitor (7Bi) in reducing Akt-pS473 signals in multiple NSCLC and HEK293 cells by blocking OTUD7B-governed G\u03b2L deubiquitination in cells, as well as inhibiting OTUD7B-mediated cleavage of K11-linked di-ub in an in vitro enzyme assay. Furthermore, we report in leukemia cells, either genetic depletion or 7Bi-mediated pharmacological inhibition of OTUD7B reduces Akt-pS473 via inhibiting the OTUD7B\/G\u03b2L signaling axis. Together, our study identifies the first putative OTUD7B inhibitor showing activities both in cells and in vitro, with promising applications as a therapeutic agent in treating cancer with OTUD7B overexpression.<\/jats:p>","DOI":"10.3390\/cancers15020517","type":"journal-article","created":{"date-parts":[[2023,1,16]],"date-time":"2023-01-16T04:31:32Z","timestamp":1673843492000},"page":"517","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["AtomNet-Aided OTUD7B Inhibitor Discovery and Validation"],"prefix":"10.3390","volume":"15","author":[{"given":"Jianfeng","family":"Chen","sequence":"first","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9062-9739","authenticated-orcid":false,"given":"Derek L.","family":"Bolhuis","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8186-0891","authenticated-orcid":false,"given":"Christian","family":"Laggner","sequence":"additional","affiliation":[{"name":"Atomwise Inc., San Francisco, CA 94103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deyu","family":"Kong","sequence":"additional","affiliation":[{"name":"Center for Integrative Chemical Biology and Drug Discovery, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Le","family":"Yu","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodong","family":"Wang","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Center for Integrative Chemical Biology and Drug Discovery, Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Emanuele","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6141-0164","authenticated-orcid":false,"given":"Nicholas G.","family":"Brown","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2755-3601","authenticated-orcid":false,"given":"Pengda","family":"Liu","sequence":"additional","affiliation":[{"name":"Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"},{"name":"Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41392-020-0107-0","article-title":"The role of ubiquitination in tumorigenesis and targeted drug discovery","volume":"5","author":"Deng","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.molcel.2021.10.027","article-title":"Deubiquitinases: From mechanisms to their inhibition by small molecules","volume":"82","author":"Lange","year":"2022","journal-title":"Mol. Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1146\/annurev-biochem-061516-044916","article-title":"Mechanisms of Deubiquitinase Specificity and Regulation","volume":"86","author":"Mevissen","year":"2017","journal-title":"Annu. Rev. Biochem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.molcel.2018.02.023","article-title":"Discovery and Characterization of ZUFSP\/ZUP1, a Distinct Deubiquitinase Class Important for Genome Stability","volume":"70","author":"Kwasna","year":"2018","journal-title":"Mol. Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.semcancer.2020.03.008","article-title":"Dissenting degradation: Deubiquitinases in cell cycle and cancer","volume":"67","author":"Bonacci","year":"2020","journal-title":"Semin. Cancer Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1038\/s41418-020-00700-z","article-title":"Deubiquitinating enzyme OTUB1 promotes cancer cell immunosuppression via preventing ER-associated degradation of immune checkpoint protein PD-L1","volume":"28","author":"Zhu","year":"2021","journal-title":"Cell Death Differ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"288","DOI":"10.15252\/emmm.201505972","article-title":"OTUB1 triggers lung cancer development by inhibiting RAS monoubiquitination","volume":"8","author":"Baietti","year":"2016","journal-title":"EMBO Mol. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s12943-014-0280-2","article-title":"OTUB1 de-ubiquitinating enzyme promotes prostate cancer cell invasion in vitro and tumorigenesis in vivo","volume":"14","author":"Chuan","year":"2015","journal-title":"Mol. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.1111\/cas.14876","article-title":"OTUB1 facilitates bladder cancer progression by stabilizing ATF6 in response to endoplasmic reticulum stress","volume":"112","author":"Zhang","year":"2021","journal-title":"Cancer Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1186\/s13578-020-00408-0","article-title":"OTUB1-mediated deubiquitination of FOXM1 up-regulates ECT-2 to promote tumor progression in renal cell carcinoma","volume":"10","author":"Zhou","year":"2020","journal-title":"Cell Biosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1158\/0008-5472.CAN-18-3037","article-title":"The Deubiquitylase OTUB1 Mediates Ferroptosis via Stabilization of SLC7A11","volume":"79","author":"Liu","year":"2019","journal-title":"Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Moree, S.E., Maneix, L., Iakova, P., Stossi, F., Sahin, E., and Catic, A. (2022). Imaging-Based Screening of Deubiquitinating Proteases Identifies Otubain-1 as a Stabilizer of c-MYC. Cancers, 14.","DOI":"10.3390\/cancers14030806"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1169","DOI":"10.1038\/ncb3218","article-title":"Deubiquitylase OTUD3 regulates PTEN stability and suppresses tumorigenesis","volume":"17","author":"Yuan","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Pu, Q., Lv, Y.R., Dong, K., Geng, W.W., and Gao, H.D. (2020). Tumor suppressor OTUD3 induces growth inhibition and apoptosis by directly deubiquitinating and stabilizing p53 in invasive breast carcinoma cells. BMC Cancer, 20.","DOI":"10.1186\/s12885-020-07069-9"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2914","DOI":"10.1038\/s41467-019-10824-7","article-title":"The deubiquitylase OTUD3 stabilizes GRP78 and promotes lung tumorigenesis","volume":"10","author":"Du","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/j.molcel.2020.06.020","article-title":"Acetylation-Dependent Deubiquitinase OTUD3 Controls MAVS Activation in Innate Antiviral Immunity","volume":"79","author":"Zhang","year":"2020","journal-title":"Mol. Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"110920","DOI":"10.1016\/j.celrep.2022.110920","article-title":"Opposing effects of deubiquitinase OTUD3 in innate immunity against RNA and DNA viruses","volume":"39","author":"Cai","year":"2022","journal-title":"Cell Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/j.cmet.2022.05.005","article-title":"Deubiquitinase OTUD3 regulates metabolism homeostasis in response to nutritional stresses","volume":"34","author":"Zhou","year":"2022","journal-title":"Cell Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1038\/nature22344","article-title":"TRAF2 and OTUD7B govern a ubiquitin-dependent switch that regulates mTORC2 signalling","volume":"545","author":"Wang","year":"2017","journal-title":"Nature"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/j.cell.2017.04.001","article-title":"AKT\/PKB Signaling: Navigating the Network","volume":"169","author":"Manning","year":"2017","journal-title":"Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e98701","DOI":"10.15252\/embj.201798701","article-title":"Cezanne\/OTUD7B is a cell cycle-regulated deubiquitinase that antagonizes the degradation of APC\/C substrates","volume":"37","author":"Bonacci","year":"2018","journal-title":"EMBO J."},{"key":"ref_22","first-page":"e2004504","article-title":"OTUD7B Deubiquitinates LSD1 to Govern Its Binding Partner Specificity, Homeostasis, and Breast Cancer Metastasis","volume":"8","author":"Gong","year":"2021","journal-title":"Adv. Sci. (Weinh.)"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1038\/s41419-021-03785-7","article-title":"OTUD7B stabilizes estrogen receptor alpha and promotes breast cancer cell proliferation","volume":"12","author":"Tang","year":"2021","journal-title":"Cell Death Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.15252\/embr.201438850","article-title":"Cezanne (OTUD7B) regulates HIF-1alpha homeostasis in a proteasome-independent manner","volume":"15","author":"Bremm","year":"2014","journal-title":"EMBO Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1038\/nature11831","article-title":"OTUD7B controls non-canonical NF-kappaB activation through deubiquitination of TRAF3","volume":"494","author":"Hu","year":"2013","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1084\/jem.20151426","article-title":"Otud7b facilitates T cell activation and inflammatory responses by regulating Zap70 ubiquitination","volume":"213","author":"Hu","year":"2016","journal-title":"J. Exp. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2288","DOI":"10.1080\/15548627.2022.2026098","article-title":"OTUD7B deubiquitinates SQSTM1\/p62 and promotes IRF3 degradation to regulate antiviral immunity","volume":"18","author":"Xie","year":"2022","journal-title":"Autophagy"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"553","DOI":"10.18632\/oncotarget.23074","article-title":"OTUD7B upregulation predicts a poor response to paclitaxel in patients with triple-negative breast cancer","volume":"9","author":"Chiu","year":"2018","journal-title":"Oncotarget"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"862","DOI":"10.3389\/fonc.2019.00862","article-title":"Upregulation of OTUD7B (Cezanne) Promotes Tumor Progression via AKT\/VEGF Pathway in Lung Squamous Carcinoma and Adenocarcinoma","volume":"9","author":"Lin","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"998","DOI":"10.7150\/jca.65835","article-title":"Deubiquitinase OTUD7B is a potential prognostic biomarker in diffuse large B-cell lymphoma","volume":"13","author":"Qiu","year":"2022","journal-title":"J. Cancer"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1016\/j.prp.2016.07.011","article-title":"OTUD7B and NIK expression in non-small cell lung cancer: Association with clinicopathological features and prognostic implications","volume":"212","author":"Zhang","year":"2016","journal-title":"Pathol. Res. Pract."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"109266","DOI":"10.1016\/j.abb.2022.109266","article-title":"OTUD7B (Cezanne) ameliorates fibrosis after myocardial infarction via FAK-ERK\/P38 MAPK signaling pathway","volume":"724","author":"Zhang","year":"2022","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Zhu, Z., Zhou, X., Du, H., Cloer, E.W., Zhang, J., Mei, L., Wang, Y., Tan, X., Hepperla, A.J., and Simon, J.M. (2022). STING Suppresses Mitochondrial VDAC2 to Govern RCC Growth Independent of Innate Immunity. Adv. Sci. (Weinh.), e2203718.","DOI":"10.1002\/advs.202203718"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.cell.2013.05.046","article-title":"OTU deubiquitinases reveal mechanisms of linkage specificity and enable ubiquitin chain restriction analysis","volume":"154","author":"Mevissen","year":"2013","journal-title":"Cell"},{"key":"ref_35","first-page":"e2004846","article-title":"SPOP and OTUD7A Control EWS-FLI1 Protein Stability to Govern Ewing Sarcoma Growth","volume":"8","author":"Su","year":"2021","journal-title":"Adv. Sci. (Weinh.)"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1038\/nature19836","article-title":"Molecular basis of Lys11-polyubiquitin specificity in the deubiquitinase Cezanne","volume":"538","author":"Mevissen","year":"2016","journal-title":"Nature"},{"key":"ref_37","unstructured":"Izhar Wallach, M.D. (2015). Abraham Heifets. AtomNet: A Deep Convolutional Neural Network for Bioactivity Prediction in Structure-based Drug Discovery. arXiv."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/S0092-8674(02)00808-5","article-title":"mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery","volume":"110","author":"Kim","year":"2002","journal-title":"Cell"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/2\/517\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:06:09Z","timestamp":1760119569000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/2\/517"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,14]]},"references-count":38,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["cancers15020517"],"URL":"https:\/\/doi.org\/10.3390\/cancers15020517","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,14]]}}}