{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T16:51:59Z","timestamp":1785775919178,"version":"3.56.0"},"reference-count":30,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T00:00:00Z","timestamp":1775088000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T00:00:00Z","timestamp":1775088000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["Finance code 001"],"award-info":[{"award-number":["Finance code 001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005942","name":"Funda\u00e7\u00e3o Butantan","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005942","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:p>\n                    Amblyomin\u2010X is a complex Kunitz\u2010type protease inhibitor from the tick\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Amblyomma sculptum<\/jats:italic>\n                    <\/jats:styled-content>\n                    , with selective pro\u2010apoptotic effects in tumor cells. While promising when applied exogenously, its large\u2010scale recombinant production is hindered by structural complexity. This study evaluated the\u00a0feasibility of expressing Amblyomin\u2010X via a suicide gene therapy approach and investigated its intracellular fate following gene delivery in human melanoma and nontumoral cell lines. Amblyomin\u2010X transcripts were equally detected in both cell types; however, protein detection was\u00a0restricted to nontumoral cells. Thus, we examined the mechanisms underlying Amblyomin\u2010X degradation in melanoma cells. Mutagenesis of\u00a0predicted ubiquitination\/SUMOylation sites failed to restore protein\u00a0detection, whereas inhibition of proteasome activity with MG132 restored Amblyomin\u2010X detection. This confirmed the involvement of a proteasome\u2010dependent mechanism that limits protein stability and consequently its cytotoxic activity when intracellularly produced in melanoma cells. These findings indicate that proteasomal degradation represents a tumor\u2010specific barrier to the intracellular expression of Amblyomin\u2010X in human cancer cells. Addressing this limitation could enable the potent activity of Amblyomin\u2010X to be harnessed through gene\u2010based anticancer strategies. Furthermore, these findings advance the understanding of how structurally complex Kunitz\u2010type proteins with unstructured regions may behave in the context of gene therapy.\n                  <\/jats:p>","DOI":"10.1002\/2211-5463.70224","type":"journal-article","created":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T16:40:16Z","timestamp":1775148016000},"page":"1602-1614","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Proteasomal degradation of intracellularly expressed Amblyomin\u2010X limits suicide gene therapy potential in melanoma cells"],"prefix":"10.1002","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5558-5953","authenticated-orcid":false,"given":"Victor Dal Posolo","family":"Cinel","sequence":"first","affiliation":[{"name":"Centre of Excellence in New Target Discovery Instituto Butantan  S\u00e3o Paulo Brazil"},{"name":"Molecular Biology Universidade Federal de S\u00e3o Paulo  Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos DeOcesano","family":"Pereira","sequence":"additional","affiliation":[{"name":"Centre of Excellence in New Target Discovery Instituto Butantan  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline Ramos Maia","family":"Lobba","sequence":"additional","affiliation":[{"name":"Centre of Excellence in New Target Discovery Instituto Butantan  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa Regina","family":"Fessel","sequence":"additional","affiliation":[{"name":"Innovation and Development Laboratory Instituto Butantan  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Marisa","family":"Chudzinski\u2010Tavassi","sequence":"additional","affiliation":[{"name":"Centre of Excellence in New Target Discovery Instituto Butantan  S\u00e3o Paulo Brazil"},{"name":"Molecular Biology Universidade Federal de S\u00e3o Paulo  Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,2]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.3322\/caac.21834"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.265"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.55"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2023.11.001"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2022.113324"},{"key":"e_1_2_11_7_1","article-title":"Suicide gene therapy for the treatment of high\u2010grade glioma: past lessons, present trends, and future prospects","volume":"2","author":"Hossain JA","year":"2020","journal-title":"Neurooncol Adv"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gendis.2022.04.004"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrdp.2015.3"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2007.12.011"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2016.07.011"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11010-016-2683-4"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10637-012-9871-1"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxicon.2010.04.019"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-45799-4"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2015.12.016"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2022.936107"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxrep.2018.11.014"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-63275-2"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi900870u"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2005.16.1016"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40265-015-0522-7"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2012.10.005"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA118.007292"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells13191670"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.3390\/biom4030862"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-017-9704-y"},{"key":"e_1_2_11_28_1","first-page":"1","article-title":"Proteotoxic crisis, the ubiquitin\u2010proteasome system, and cancer therapy","volume":"12","author":"Deshaies RJ","year":"2014","journal-title":"BMC Med"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-005-1581-0"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1080\/15384047.2021.1978785"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1158\/1535-7163.MCT-17-0919"}],"container-title":["FEBS Open Bio"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/2211-5463.70224","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/2211-5463.70224","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/2211-5463.70224","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T16:21:11Z","timestamp":1785774071000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1002\/2211-5463.70224"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,2]]},"references-count":30,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2026,8]]}},"alternative-id":["10.1002\/2211-5463.70224"],"URL":"https:\/\/doi.org\/10.1002\/2211-5463.70224","archive":["Portico"],"relation":{},"ISSN":["2211-5463","2211-5463"],"issn-type":[{"value":"2211-5463","type":"print"},{"value":"2211-5463","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,2]]},"assertion":[{"value":"2025-08-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-02-23","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-04-02","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}