{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T11:04:44Z","timestamp":1789383884020,"version":"build-2803163510"},"reference-count":38,"publisher":"Bioscientifica","issue":"2","content-domain":{"domain":["journals.bioscientifica.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Everolimus, an mTORC1 inhibitor, is approved for treating pancreatic and lung neuroendocrine tumors (NETs), yet acquired resistance limits its long-term efficacy. The molecular mechanisms underlying resistance and potential strategies to overcome it remain poorly defined. RNA sequencing was performed in everolimus-sensitive and everolimus-resistant QGP-1 pancreatic NET cells (QS and QR, respectively) to identify transcriptional changes associated with everolimus resistance. Key differentially expressed genes were validated in QGP-1, H727 lung NET cells, and patient-derived Pa-NET cells using RT-qPCR. The potential functional involvement of caspase-8 (CASP8) was investigated pharmacologically using the preferential CASP8 inhibitor Z-IETD-FMK, alone or in combination with everolimus, by evaluating effects on cell viability and CASP activity. Differential expression analysis identified 292 genes significantly dysregulated in QR versus QS cells, including CASP8, A2M, MGP, PTGS1, and IGFBP5. Enrichment analysis highlighted pathways related to ECM remodeling, neuroactive ligand\u2013receptor interactions, and GPCR signaling. Validation in H727 cells and primary Pa-NET samples showed some conserved transcriptional changes, with CASP8 upregulation emerging as a particularly consistent alteration. CASP8 inhibition reduced viability in both QS and QR cells, while combined Z-IETD-FMK and everolimus treatment produced the greatest decrease in QS viability and was associated with reduced CASP3\/7 activity. Our study identifies CASP8 as a candidate mediator associated with everolimus resistance in NETs, together with alterations in ECM remodeling and receptor signaling. These findings support further investigation of CASP8-targeting strategies, particularly in combination with mTOR inhibition, as a potential approach to enhance therapeutic efficacy.<\/jats:p>","DOI":"10.1530\/jme-25-0206","type":"journal-article","created":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T10:01:56Z","timestamp":1787133716000},"update-policy":"https:\/\/doi.org\/10.1530\/crossmarkpolicy-6","source":"Crossref","is-referenced-by-count":0,"title":["Caspase-8 as a candidate mediator associated with everolimus resistance in neuroendocrine tumors"],"prefix":"10.1530","volume":"77","author":[{"given":"Cristina","family":"Grassi","sequence":"first","affiliation":[{"name":"Laboratory of Translational Endocrinology and Metabolism, IRCCS Humanitas Research Hospital 1 , Rozzano, ,","place":["Milan, Italy"]},{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam","family":"Bondi","sequence":"additional","affiliation":[{"name":"Laboratory of Translational Endocrinology and Metabolism, IRCCS Humanitas Research Hospital 1 , Rozzano, ,","place":["Milan, Italy"]},{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giulia","family":"Albanese","sequence":"additional","affiliation":[{"name":"Laboratory of Translational Endocrinology and Metabolism, IRCCS Humanitas Research Hospital 1 , Rozzano, ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Zerbi","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]},{"name":"Pancreatic Surgery Unit, IRCCS Humanitas Research Hospital 3 ,","place":["Rozzano, Milan,, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Uccella","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]},{"name":"Pathology Unit, IRCCS Humanitas Research Hospital 4 , , Milan,","place":["Rozzano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4424-7895","authenticated-orcid":true,"given":"Erika","family":"Peverelli","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences and Community Health, Department of Excellence 2023\u20132027, University of Milan 5 , ,","place":["Milan, Italy"]},{"name":"Endocrinology Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico 6 , ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanna","family":"Mantovani","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences and Community Health, Department of Excellence 2023\u20132027, University of Milan 5 , ,","place":["Milan, Italy"]},{"name":"Endocrinology Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico 6 , ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabetta","family":"Lavezzi","sequence":"additional","affiliation":[{"name":"Endocrinology, Diabetology and Andrology Unit, IRCCS Humanitas Research Hospital 7 , , Milan,","place":["Rozzano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Marulli","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]},{"name":"Thoracic Surgery Unit, IRCCS Humanitas Research Hospital 8 , , Milan,","place":["Rozzano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea Gerardo Antonio","family":"Lania","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Humanitas University 2 , Pieve Emanuele, ,","place":["Milan, Italy"]},{"name":"Endocrinology, Diabetology and Andrology Unit, IRCCS Humanitas Research Hospital 7 , , Milan,","place":["Rozzano, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1827-0530","authenticated-orcid":true,"given":"Eleonora","family":"Vitali","sequence":"additional","affiliation":[{"name":"Laboratory of Translational Endocrinology and Metabolism, IRCCS Humanitas Research Hospital 1 , Rozzano, ,","place":["Milan, Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"416","published-online":{"date-parts":[[2026,8,19]]},"reference":[{"key":"2026091406314219300_bib1","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1016\/j.neo.2017.09.002","article-title":"Nothing but NET: a review of neuroendocrine tumors and carcinomas","volume":"19","author":"Oronsky","year":"2017","journal-title":"Neoplasia"},{"key":"2026091406314219300_bib2","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1016\/j.hlife.2024.07.006","article-title":"Current perspectives on neuroendocrine tumors","volume":"2","author":"Verma","year":"2024","journal-title":"hLife"},{"key":"2026091406314219300_bib3","doi-asserted-by":"crossref","first-page":"3794","DOI":"10.3390\/jcm11133794","article-title":"The increasing incidence of neuroendocrine neoplasms worldwide: current knowledge and open issues","volume":"11","author":"Rossi","year":"2022","journal-title":"J Clin Med"},{"key":"2026091406314219300_bib4","doi-asserted-by":"crossref","DOI":"10.1007\/978-3-031-56968-5","volume-title":"Neuroendocrine Tumours: Diagnosis and Management","author":"Yalcin","year":"2024","edition":"2nd ed"},{"key":"2026091406314219300_bib5","doi-asserted-by":"crossref","first-page":"3090","DOI":"10.1002\/cncr.35463","article-title":"Update in the management of gastroenteropancreatic neuroendocrine tumors","volume":"130","author":"Sedlack","year":"2024","journal-title":"Cancer"},{"key":"2026091406314219300_bib6","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1007\/s12022-022-09708-2","article-title":"Overview of the 2022 WHO classification of neuroendocrine neoplasms","volume":"33","author":"Rindi","year":"2022","journal-title":"Endocr Pathol"},{"key":"2026091406314219300_bib7","article-title":"Differentiation between G3 pancreatic neuroendocrine tumor and pancreatic neuroendocrine carcinoma based on intratumor and peritumor CT value ratio and abnormal vascular network","volume":"15","author":"Zhang","year":"2025","journal-title":"Front Oncol"},{"key":"2026091406314219300_bib8","doi-asserted-by":"crossref","first-page":"828","DOI":"10.3390\/cancers11060828","article-title":"Treatment options for pancreatic neuroendocrine tumors","volume":"11","author":"Akirov","year":"2019","journal-title":"Cancers"},{"key":"2026091406314219300_bib9","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1016\/j.annonc.2020.03.304","article-title":"Gastroenteropancreatic neuroendocrine neoplasms: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up","volume":"31","author":"Pavel","year":"2020","journal-title":"Ann Oncol"},{"key":"2026091406314219300_bib10","doi-asserted-by":"crossref","DOI":"10.1016\/j.currproblcancer.2024.101061","article-title":"Current and emerging strategies for the management of advanced\/metastatic lung neuroendocrine tumors","volume":"49","author":"Rutherford","year":"2024","journal-title":"Curr Probl Cancer"},{"key":"2026091406314219300_bib11","doi-asserted-by":"crossref","first-page":"640","DOI":"10.1056\/NEJMoa1916623","article-title":"The effect of advances in lung-cancer treatment on population mortality","volume":"383","author":"Howlader","year":"2020","journal-title":"N Engl J Med"},{"key":"2026091406314219300_bib12","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1159\/000443167","article-title":"ENETS consensus guidelines update for the management of distant metastatic disease of intestinal, pancreatic, bronchial neuroendocrine neoplasms (NEN) and NEN of unknown primary site","volume":"103","author":"Pavel","year":"2016","journal-title":"Neuroendocrinology"},{"key":"2026091406314219300_bib13","doi-asserted-by":"crossref","DOI":"10.3389\/fendo.2020.562505","article-title":"mTOR pathway in gastroenteropancreatic neuroendocrine tumor (GEP-NETs)","volume":"11","author":"Zanini","year":"2020","journal-title":"Front Endocrinol"},{"key":"2026091406314219300_bib14","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1016\/S1470-2045(17)30471-0","article-title":"Health-related quality of life for everolimus versus placebo in patients with advanced, non-functional, well-differentiated gastrointestinal or lung neuroendocrine tumours (RADIANT-4): a multicentre, randomised, double-blind, placebo-controlled, phase 3 trial","volume":"18","author":"Pavel","year":"2017","journal-title":"Lancet Oncol"},{"key":"2026091406314219300_bib15","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1056\/NEJMoa1009290","article-title":"Everolimus for advanced pancreatic neuroendocrine tumors for the RAD001 in advanced neuroendocrine tumors, third trial (RADIANT-3) study group","volume":"364","author":"Yao","year":"2011","journal-title":"N Engl J Med"},{"key":"2026091406314219300_bib16","doi-asserted-by":"crossref","first-page":"455","DOI":"10.3390\/biomedicines13020455","article-title":"Everolimus in the treatment of neuroendocrine tumors: lights and shadows","volume":"13","author":"Medici","year":"2025","journal-title":"Biomedicines"},{"key":"2026091406314219300_bib17","doi-asserted-by":"crossref","first-page":"7409","DOI":"10.1158\/0008-5472.CAN-08-1522","article-title":"Enhancing mammalian target of rapamycin (mTOR)-targeted cancer therapy by preventing mTOR\/raptor inhibition-initiated, mTOR\/rictor-independent Akt activation","volume":"68","author":"Wang","year":"2008","journal-title":"Cancer Res"},{"key":"2026091406314219300_bib18","doi-asserted-by":"crossref","first-page":"2587","DOI":"10.3390\/ijms25052587","article-title":"mTOR signaling: recent progress","volume":"25","author":"Gargalionis","year":"2024","journal-title":"Int J Mol Sci"},{"key":"2026091406314219300_bib19","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1007\/s40618-023-02221-1","article-title":"Pancreatic neuroendocrine tumor progression and resistance to everolimus: the crucial role of NF-kB and STAT3 interplay","volume":"47","author":"Vitali","year":"2023","journal-title":"J Endocrinol Investig"},{"key":"2026091406314219300_bib20","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1080\/14656566.2018.1476492","article-title":"Everolimus in the treatment of neuroendocrine tumors: efficacy, side-effects, resistance, and factors affecting its place in the treatment sequence","volume":"19","author":"Lee","year":"2018","journal-title":"Expert Opin Pharmacother"},{"key":"2026091406314219300_bib21","doi-asserted-by":"crossref","first-page":"S89","DOI":"10.1016\/j.ijsu.2015.06.064","article-title":"Everolimus and pancreatic neuroendocrine tumors (PNETs): activity, resistance and how to overcome it","volume":"21","author":"Capozzi","year":"2015","journal-title":"Int J Surg"},{"key":"2026091406314219300_bib22","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1038\/bjc.2016.25","article-title":"Long-term acquired everolimus resistance in pancreatic neuroendocrine tumours can be overcome with novel PI3K-AKT-mTOR inhibitors","volume":"114","author":"Vandamme","year":"2016","journal-title":"Br J Cancer"},{"key":"2026091406314219300_bib23","doi-asserted-by":"crossref","first-page":"2249","DOI":"10.3390\/cancers15082249","article-title":"Neuroendocrine tumors: genomics and molecular biomarkers with a focus on metastatic disease","volume":"15","author":"Alexander","year":"2023","journal-title":"Cancers"},{"key":"2026091406314219300_bib24","doi-asserted-by":"crossref","DOI":"10.1016\/j.mce.2020.110803","article-title":"A novel insight into the anticancer mechanism of metformin in pancreatic neuroendocrine tumor cells","volume":"509","author":"Vitali","year":"2020","journal-title":"Mol Cell Endocrinol"},{"key":"2026091406314219300_bib25","doi-asserted-by":"crossref","first-page":"2143","DOI":"10.3390\/cancers12082143","article-title":"Metformin and everolimus: a promising combination for neuroendocrine tumors treatment","volume":"12","author":"Vitali","year":"2020","journal-title":"Cancers (Basel)"},{"key":"2026091406314219300_bib26","volume-title":"Neuroendocrine Neoplasia","author":"La Salvia","year":"2025"},{"key":"2026091406314219300_bib27","doi-asserted-by":"crossref","DOI":"10.1007\/s11912-021-01029-7","article-title":"Epidemiology, incidence, and prevalence of neuroendocrine neoplasms: are there global differences?","volume":"23","author":"Das","year":"2021","journal-title":"Curr Oncol Rep"},{"key":"2026091406314219300_bib28","doi-asserted-by":"crossref","first-page":"R109","DOI":"10.1530\/ERC-18-0420","article-title":"Resistance to targeted treatment of gastroenteropancreatic neuroendocrine tumors","volume":"26","author":"Beyens","year":"2019","journal-title":"Endocr Relat Cancer"},{"key":"2026091406314219300_bib29","doi-asserted-by":"crossref","first-page":"3271","DOI":"10.3390\/cancers15133271","article-title":"Caspase-8 and tyrosine kinases: a dangerous liaison in cancer","volume":"15","author":"Contadini","year":"2023","journal-title":"Cancers"},{"key":"2026091406314219300_bib30","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1002\/ijc.33698","article-title":"Caspase-8: a key protein of cross-talk signal way in \u2018PANoptosis\u2019 in cancer","volume":"149","author":"Jiang","year":"2021","journal-title":"Int J Cancer"},{"key":"2026091406314219300_bib31","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1038\/s41419-021-04126-4","article-title":"Non-apoptotic function of caspase-8 confers prostate cancer enzalutamide resistance via NF-\u03baB activation","volume":"12","author":"Xia","year":"2021","journal-title":"Cell Death Dis"},{"key":"2026091406314219300_bib32","doi-asserted-by":"crossref","first-page":"3798","DOI":"10.3390\/ijms19123798","article-title":"Caspase-8: a novel target to overcome resistance to chemotherapy in glioblastoma","volume":"19","author":"Fianco","year":"2018","journal-title":"Int J Mol Sci"},{"key":"2026091406314219300_bib33","doi-asserted-by":"crossref","DOI":"10.1016\/j.ejca.2025.115267","article-title":"The role of IL-8 in cancer development and its impact on immunotherapy resistance","volume":"218","author":"Meier","year":"2025","journal-title":"Eur J Cancer"},{"key":"2026091406314219300_bib34","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.semcdb.2018.01.009","article-title":"Caspase-8 function, and phosphorylation, in cell migration","volume":"82","author":"Keller","year":"2018","journal-title":"Semin Cell Dev Biol"},{"key":"2026091406314219300_bib35","doi-asserted-by":"crossref","DOI":"10.1016\/j.bbcan.2020.188357","article-title":"Caspase-8: the double-edged sword","volume":"1873","author":"Mandal","year":"2020","journal-title":"Biochim Biophys Acta - Rev Cancer"},{"key":"2026091406314219300_bib36","doi-asserted-by":"crossref","first-page":"5805","DOI":"10.1007\/s12035-023-03433-5","article-title":"A review on caspases: key regulators of biological activities and apoptosis","volume":"60","author":"Sahoo","year":"2023","journal-title":"Mol Neurobiol"},{"key":"2026091406314219300_bib37","doi-asserted-by":"crossref","DOI":"10.3389\/fonc.2020.01563","article-title":"Matrix stiffness-upregulated MicroRNA-17-5p attenuates the intervention effects of metformin on HCC invasion and metastasis by targeting the PTEN\/PI3K\/Akt pathway","volume":"10","author":"Gao","year":"2020","journal-title":"Front Oncol"},{"key":"2026091406314219300_bib38","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1186\/s40164-025-00647-2","article-title":"Extracellular matrix stiffness: mechanisms in tumor progression and therapeutic potential in cancer","volume":"14","author":"Zhang","year":"2025","journal-title":"Exp Hematol Oncol"}],"container-title":["Journal of Molecular Endocrinology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.bioscientifica.com\/jme\/article-pdf\/doi\/10.1530\/JME-25-0206\/524023\/jme-25-0206.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.bioscientifica.com\/jme\/article-pdf\/doi\/10.1530\/JME-25-0206\/524023\/jme-25-0206.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T10:31:51Z","timestamp":1789381911000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.bioscientifica.com\/jme\/article\/77\/2\/e250206\/78055\/Caspase-8-as-a-candidate-mediator-associated-with"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,1]]},"references-count":38,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2026,8,1]]}},"URL":"https:\/\/doi.org\/10.1530\/jme-25-0206","relation":{},"ISSN":["0952-5041","1479-6813"],"issn-type":[{"value":"0952-5041","type":"print"},{"value":"1479-6813","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,1]]},"article-number":"e250206"}}