{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:35:43Z","timestamp":1760060143079,"version":"build-2065373602"},"reference-count":41,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,8,8]],"date-time":"2025-08-08T00:00:00Z","timestamp":1754611200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"AIRC (Italian Association for Cancer Research)","award":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"],"award-info":[{"award-number":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"]}]},{"name":"Italian Ministry of Research","award":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"],"award-info":[{"award-number":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"]}]},{"name":"National Center for Gene Therapy and Drug based on RNA Technology","award":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"],"award-info":[{"award-number":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"]}]},{"name":"NextGenerationEU PNRR MUR-M4C2-Action 1.4","award":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"],"award-info":[{"award-number":["IG 21803","23560","PRIN2017\u201420174PLLYN_004","CN00000041"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>The tumor suppressor DAB2IP, a RasGAP and cytoplasmic adaptor protein, modulates signal transduction in response to several extracellular stimuli, negatively regulating multiple oncogenic pathways. Accordingly, the loss of DAB2IP in tumor cells fosters metastasis and enhances chemo- and radioresistance. DAB2IP is rarely mutated in cancer but is frequently downregulated or inactivated by multiple mechanisms. Solid experimental evidence shows that DAB2IP reactivation reduces cancer aggressiveness in tumors driven by multiple different oncogenic mutations, making this protein an interesting target for cancer therapy. Considering this evidence, we screened a drug library to identify molecules that increase DAB2IP protein levels. We employed CRISPR\/Cas9 gene editing to generate two prostate cancer cell models in which endogenous DAB2IP is fused to HiBiT, a peptide tag that enables luminescence-based detection of protein levels in a sensitive and quantitative manner. Using this approach, we identified drugs able to increase DAB2IP levels. We focused our attention on thiostrepton, a natural cyclic oligopeptide antibiotic that has been reported to inhibit the survival of various cancer cell lines. Functional experiments revealed that the cancer-inhibitory effect of thiostrepton is reduced in the absence of DAB2IP, suggesting that upregulation of this protein contributes to its action. These findings encourage further development of thiostrepton for the treatment of solid cancers and unveil a novel molecular target underlying its anti-tumoral activity.<\/jats:p>","DOI":"10.3390\/biom15081147","type":"journal-article","created":{"date-parts":[[2025,8,8]],"date-time":"2025-08-08T13:20:09Z","timestamp":1754659209000},"page":"1147","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Drug-Repurposing Screen Identifies Thiostrepton as a Novel Regulator of the Tumor Suppressor DAB2IP"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-4731-2479","authenticated-orcid":false,"given":"Rossella","family":"De Florian Fania","sequence":"first","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via L. Giorgieri 1, 34127 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Serena","family":"Maiocchi","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via L. Giorgieri 1, 34127 Trieste, Italy"},{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano 99, 34149 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raffaella","family":"Klima","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano 99, 34149 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monica","family":"Rossin","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via L. Giorgieri 1, 34127 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valeria","family":"Pellegrini","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via L. Giorgieri 1, 34127 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4981-9012","authenticated-orcid":false,"given":"Sabrina","family":"Ghetti","sequence":"additional","affiliation":[{"name":"Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Via Sommarive 9, 38123 Povo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Davide","family":"Selvestrel","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano 99, 34149 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Chiara","family":"Mattevi","sequence":"additional","affiliation":[{"name":"Unit of Molecular Oncology, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, National Cancer Institute, Via Gallini 2, 33081 Aviano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca L.","family":"Fava","sequence":"additional","affiliation":[{"name":"Armenise-Harvard Laboratory of Cell Division, Department of Cellular, Computational and Integrative Biology (CIBIO), University of Trento, Via Sommarive 9, 38123 Povo, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9827-6755","authenticated-orcid":false,"given":"Luca","family":"Braga","sequence":"additional","affiliation":[{"name":"International Centre for Genetic Engineering and Biotechnology (ICGEB), Padriciano 99, 34149 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6815-5381","authenticated-orcid":false,"given":"Licio","family":"Collavin","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Trieste, Via L. Giorgieri 1, 34127 Trieste, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"63","DOI":"10.14740\/wjon1191","article-title":"Epidemiology of prostate cancer","volume":"10","author":"Rawla","year":"2019","journal-title":"World J. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1038\/cdd.2016.134","article-title":"Block one, unleash a hundred. Mechanisms of DAB2IP inactivation in cancer","volume":"24","author":"Bellazzo","year":"2017","journal-title":"Cell Death Differ."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3766","DOI":"10.18632\/oncotarget.6501","article-title":"DAB2IP in cancer","volume":"7","author":"Liu","year":"2016","journal-title":"Oncotarget"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1038\/s41418-024-01332-3","article-title":"An update on the tumor-suppressive functions of the RasGAP protein DAB2IP with focus on therapeutic implications","volume":"31","author":"Bellazzo","year":"2024","journal-title":"Cell Death Differ."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1933","DOI":"10.1172\/JCI200317790","article-title":"AIP1 mediates TNF-alpha-induced ASK1 activation by facilitating dissociation of ASK1 from its inhibitor 14-3-3","volume":"111","author":"Zhang","year":"2003","journal-title":"J. Clin. Investig."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Bellazzo, A., and Collavin, L. (2020). Cutting the Brakes on Ras\u2014Cytoplasmic GAPs as Targets of Inactivation in Cancer. Cancers, 12.","DOI":"10.3390\/cancers12103066"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1158\/2159-8290.CD-16-0520","article-title":"Loss of RasGAP Tumor Suppressors Underlies the Aggressive Nature of Luminal B Breast Cancers","volume":"7","author":"Olsen","year":"2017","journal-title":"Cancer Discov."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1038\/s41418-018-0088-5","article-title":"Cell-autonomous and cell non-autonomous downregulation of tumor suppressor DAB2IP by microRNA-149-3p promotes aggressiveness of cancer cells","volume":"25","author":"Bellazzo","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1016\/j.molcel.2014.10.013","article-title":"Mutant p53 reprograms TNF signaling in cancer cells through interaction with the tumor suppressor DAB2IP","volume":"56","author":"Bellazzo","year":"2014","journal-title":"Mol. Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"7623","DOI":"10.1073\/pnas.1700996114","article-title":"Mutant p53 potentiates the oncogenic effects of insulin by inhibiting the tumor suppressor DAB2IP","volume":"114","author":"Valentino","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Schwinn, M.K., Steffen, L.S., Zimmerman, K., Wood, K.V., and Machleidt, T. (2020). A Simple and Scalable Strategy for Analysis of Endogenous Protein Dynamics. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-65832-1"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"103000","DOI":"10.1016\/j.xpro.2024.103000","article-title":"Protocol for HiBiT tagging endogenous proteins using CRISPR-Cas9 gene editing","volume":"5","author":"Lankford","year":"2024","journal-title":"STAR Protoc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"100407","DOI":"10.1016\/j.xpro.2021.100407","article-title":"CRISPR\/Cas9 ribonucleoprotein-mediated knockin generation in hTERT-RPE1 cells","volume":"2","author":"Ghetti","year":"2021","journal-title":"STAR Protoc."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Mittler, F., Obeid, P., Rulina, A.V., Haguet, V., Gidrol, X., and Balakirev, M.Y. (2017). High-Content Monitoring of Drug Effects in a 3D Spheroid Model. Front. Oncol., 7.","DOI":"10.3389\/fonc.2017.00293"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1038\/nm.2100","article-title":"An oncogene-tumor suppressor cascade drives metastatic prostate cancer by coordinately activating Ras and nuclear factor-kappaB","volume":"16","author":"Min","year":"2010","journal-title":"Nat. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4740","DOI":"10.1158\/1078-0432.CCR-13-0954","article-title":"The mechanism of DAB2IP in chemoresistance of prostate cancer cells","volume":"19","author":"Wu","year":"2013","journal-title":"Clin. Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"174661","DOI":"10.1016\/j.ejphar.2021.174661","article-title":"The bacterial thiopeptide thiostrepton. An update of its mode of action, pharmacological properties and applications","volume":"914","author":"Bailly","year":"2022","journal-title":"Eur. J. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"964","DOI":"10.2174\/092986711794940879","article-title":"Progress in the development of bestatin analogues as aminopeptidases inhibitors","volume":"18","author":"Chen","year":"2011","journal-title":"Curr. Med. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Barnieh, F.M., Loadman, P.M., and Falconer, R.A. (2021). Is tumour-expressed aminopeptidase N (APN\/CD13) structurally and functionally unique?. Biochim. Biophys. Acta Rev. Cancer, 1876.","DOI":"10.1016\/j.bbcan.2021.188641"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"19","DOI":"10.2165\/00003088-200241010-00003","article-title":"Clinical pharmacokinetics of salmeterol","volume":"41","author":"Cazzola","year":"2002","journal-title":"Clin. Pharmacokinet."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1111\/j.1476-5381.2009.00296.x","article-title":"Molecular mechanisms for the persistent bronchodilatory effect of the beta 2-adrenoceptor agonist salmeterol","volume":"158","author":"Szczuka","year":"2009","journal-title":"Br. J. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/j.freeradbiomed.2022.09.018","article-title":"Thiostrepton confers protection against reactive oxygen species-related apoptosis by restraining FOXM1-triggerred development of gastric cancer","volume":"193","author":"Liu","year":"2022","journal-title":"Free Radic. Biol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2022","DOI":"10.1158\/1535-7163.MCT-08-0188","article-title":"Thiostrepton selectively targets breast cancer cells through inhibition of forkhead box M1 expression","volume":"7","author":"Kwok","year":"2008","journal-title":"Mol. Cancer Ther."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Kalathil, D., John, S., and Nair, A.S. (2020). FOXM1 and Cancer: Faulty Cellular Signaling Derails Homeostasis. Front. Oncol., 10.","DOI":"10.3389\/fonc.2020.626836"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1038\/s41419-017-0156-7","article-title":"Honokiol is a FOXM1 antagonist","volume":"9","author":"Halasi","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5165","DOI":"10.1038\/ncomms6165","article-title":"Suppression of the FOXM1 transcriptional programme via novel small molecule inhibition","volume":"5","author":"Gormally","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1038\/s41587-022-01539-0","article-title":"A proteome-wide atlas of drug mechanism of action","volume":"41","author":"Mitchell","year":"2023","journal-title":"Nat. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6425","DOI":"10.18632\/oncotarget.2228","article-title":"The role of homeostatic regulation between tumor suppressor DAB2IP and oncogenic Skp2 in prostate cancer growth","volume":"5","author":"Tsai","year":"2014","journal-title":"Oncotarget"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"26057","DOI":"10.18632\/oncotarget.8424","article-title":"Smurf1 regulation of DAB2IP controls cell proliferation and migration","volume":"7","author":"Li","year":"2016","journal-title":"Oncotarget"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1016\/j.febslet.2015.03.027","article-title":"miR-889 promotes proliferation of esophageal squamous cell carcinomas through DAB2IP","volume":"589","author":"Xu","year":"2015","journal-title":"FEBS Lett."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e12630","DOI":"10.1111\/cpr.12630","article-title":"miR-92b promotes gastric cancer growth by activating the DAB2IP-mediated PI3K\/AKT signalling pathway","volume":"53","author":"Ni","year":"2020","journal-title":"Cell Prolif."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.biocel.2019.04.002","article-title":"miR-182 contributes to cell proliferation, invasion and tumor growth in colorectal cancer by targeting DAB2IP","volume":"111","author":"Li","year":"2019","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2287","DOI":"10.1158\/1535-7163.MCT-11-0536","article-title":"Micelle-encapsulated thiostrepton as an effective nanomedicine for inhibiting tumor growth and for suppressing FOXM1 in human xenografts","volume":"10","author":"Wang","year":"2011","journal-title":"Mol. Cancer Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1038\/s41419-022-05082-3","article-title":"Thiostrepton induces ferroptosis in pancreatic cancer cells through STAT3\/GPX4 signalling","volume":"13","author":"Zhang","year":"2022","journal-title":"Cell Death Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1801","DOI":"10.1038\/cddis.2015.155","article-title":"Identification of thiostrepton as a novel therapeutic agent that targets human colon cancer stem cells","volume":"6","author":"Ju","year":"2015","journal-title":"Cell Death Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1007\/s13402-021-00603-2","article-title":"Identification of lysosome-targeting drugs with anti-inflammatory activity as potential invasion inhibitors of treatment resistant HER2 positive cancers","volume":"44","author":"Hansen","year":"2021","journal-title":"Cell. Oncol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1007\/s00432-014-1872-3","article-title":"Targeting FoxM1 by thiostrepton inhibits growth and induces apoptosis of laryngeal squamous cell carcinoma","volume":"141","author":"Jiang","year":"2015","journal-title":"J. Cancer. Res. Clin. Oncol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1016\/j.bcp.2012.01.027","article-title":"Thiostrepton is an inducer of oxidative and proteotoxic stress that impairs viability of human melanoma cells but not primary melanocytes","volume":"83","author":"Qiao","year":"2012","journal-title":"Biochem. Pharmacol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Ismail, T., Kim, Y., Lee, H., Lee, D.-S., and Lee, H.-S. (2019). Interplay Between Mitochondrial Peroxiredoxins and ROS in Cancer Development and Progression. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20184407"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Nelson, K.J., Messier, T., Milczarek, S., Saaman, A., Beuschel, S., Gandhi, U., Heintz, N., Smalley, T.L., Lowther, W.T., and Cunniff, B. (2021). Unique Cellular and Biochemical Features of Human Mitochondrial Peroxiredoxin 3 Establish the Molecular Basis for Its Specific Reaction with Thiostrepton. Antioxidants, 10.","DOI":"10.3390\/antiox10020150"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Cunniff, B., Newick, K., Nelson, K.J., Wozniak, A.N., Beuschel, S., Leavitt, B., Bhave, A., Butnor, K., Koenig, A., and Chouchani, E.T. (2015). Disabling Mitochondrial Peroxide Metabolism via Combinatorial Targeting of Peroxiredoxin 3 as an Effective Therapeutic Approach for Malignant Mesothelioma. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127310"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/8\/1147\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:26:35Z","timestamp":1760034395000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/8\/1147"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,8]]},"references-count":41,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["biom15081147"],"URL":"https:\/\/doi.org\/10.3390\/biom15081147","relation":{},"ISSN":["2218-273X"],"issn-type":[{"type":"electronic","value":"2218-273X"}],"subject":[],"published":{"date-parts":[[2025,8,8]]}}}