{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T20:29:21Z","timestamp":1773260961074,"version":"3.50.1"},"reference-count":53,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,11]],"date-time":"2026-01-11T00:00:00Z","timestamp":1768089600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"SECIHTI","award":["CBF-2025-1-4446"],"award-info":[{"award-number":["CBF-2025-1-4446"]}]},{"name":"the Instituto Nacional de Medicina Gen\u00f3mica"},{"name":"SECIHTI","award":["CVU.781517"],"award-info":[{"award-number":["CVU.781517"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The aggressiveness of pancreatic ductal adenocarcinoma (PDAC) has been linked to cancer stem cells (CSCs) and telomerase activity; however, the mechanism underlying this association remains unclear. In this study, we engineered dual transcriptional reporters (SORE6-GFP and TERT-BFP) to isolate SOX2+OCT4+TERThigh subpopulations from AsPC-1 and BxPC-3 cells. We combined Fluorescence-Activated Cell Sorting with functional assays, RNA-seq, and network analysis. Clinically, tumors co-expressing high SOX2\/OCT4\/TERT levels were associated with reduced overall survival, whereas single-gene elevations were not prognostic. We identified a minority SOX2+OCT4+TERThigh fraction (~9%) enriched for pluripotency transcripts (SOX2, OCT4, NANOG, and ALDH1A1), which exhibited the highest proliferative, migratory, and invasive capacities. Transcriptomic profiling of SOX2+OCT4+TERThigh cells showed enrichment of KRAS, telomere maintenance, epithelial\u2013mesenchymal transition, and developmental pathways (WNT and Hedgehog). Connectivity profiling highlighted actionable vulnerabilities, including NF-\u03baB, WNT, and telomerase inhibition pathways. Together, these data define an aggressive telomerase-engaged, pluripotency-driven CSC-like state in PDAC and suggest testable therapeutic strategies that target TERThigh dependencies.<\/jats:p>","DOI":"10.3390\/cells15020129","type":"journal-article","created":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T07:30:34Z","timestamp":1768203034000},"page":"129","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Pancreatic Cancer Stem Cells Co-Expressing SOX2, OCT4, and TERThigh Represent an Aggressive Subpopulation"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-9686-9065","authenticated-orcid":false,"given":"Erika","family":"Curiel-Gomez","sequence":"first","affiliation":[{"name":"Posgrado en Ciencias Biol\u00f3gicas, Unidad de Posgrado, Edificio D, 1\u00b0 Piso, Circuito de Posgrados, Ciudad Universitaria, Coyoac\u00e1n, Mexico City C.P. 04510, Mexico"},{"name":"Laboratorio de Geno\u0301mica Funcional del Ca\u0301ncer, Instituto Nacional de Medicina Gen\u00f3mica, Perif\u00e9rico Sur No. 4809, Col Arenal Tepepan, Tlalpan, Mexico City C.P. 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Damaris P.","family":"Romero-Rodriguez","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional Conahcyt de Investigaci\u00f3n y Diagn\u00f3stico por Inmunocitofluorometr\u00eda (LANCIDI), Instituto Nacional de Enfermedades Respiratorias \u201cIsmael Cos\u00edo Villegas\u201d, Calzada de Tlalpan 4502, Col. Belisario Dom\u00ednguez Secc. 16, Tlalpan, Mexico City C.P. 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4249-7222","authenticated-orcid":false,"given":"Mauricio","family":"Rodriguez-Dorantes","sequence":"additional","affiliation":[{"name":"Laboratorio de Oncogen\u00f3mica, Instituto Nacional de Medicina Gen\u00f3mica, Perif\u00e9rico Sur No. 4809, Col Arenal Tepepan, Tlalpan, Mexico City C.P. 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7254-5961","authenticated-orcid":false,"given":"Vilma","family":"Maldonado","sequence":"additional","affiliation":[{"name":"Laboratorio de Epigene\u0301tica, Instituto Nacional de Medicina Gen\u00f3mica, Perif\u00e9rico Sur No. 4809, Col Arenal Tepepan, Tlalpan, Mexico City C.P. 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2209-1607","authenticated-orcid":false,"given":"Jorge","family":"Melendez-Zajgla","sequence":"additional","affiliation":[{"name":"Laboratorio de Geno\u0301mica Funcional del Ca\u0301ncer, Instituto Nacional de Medicina Gen\u00f3mica, Perif\u00e9rico Sur No. 4809, Col Arenal Tepepan, Tlalpan, Mexico City C.P. 14610, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,11]]},"reference":[{"key":"ref_1","first-page":"17","article-title":"Cancer statistics, 2023","volume":"73","author":"Siegel","year":"2023","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e214708","DOI":"10.1001\/jamanetworkopen.2021.4708","article-title":"Estimated Projection of US Cancer Incidence and Death to 2040","volume":"4","author":"Rahib","year":"2021","journal-title":"JAMA Netw. Open"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1729","DOI":"10.1016\/j.cell.2023.02.014","article-title":"Pancreatic Cancer: Advances and Challenges","volume":"186","author":"Halbrook","year":"2023","journal-title":"Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1817","DOI":"10.1056\/NEJMoa1011923","article-title":"FOLFIRINOX versus Gemcitabine for Metastatic Pancreatic Cancer","volume":"364","author":"Conroy","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1691","DOI":"10.1056\/NEJMoa1304369","article-title":"Increased Survival in Pancreatic Cancer with nab-Paclitaxel plus Gemcitabine","volume":"369","author":"Hoff","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.stem.2007.06.002","article-title":"Distinct Populations of Cancer Stem Cells Determine Tumor Growth and Metastatic Activity in Human Pancreatic Cancer","volume":"1","author":"Hermann","year":"2007","journal-title":"Cell Stem Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1390","DOI":"10.1038\/s41588-022-01157-1","article-title":"Spatially Restricted Drivers and Transitional Cell Populations Cooperate with the Microenvironment in Untreated and Chemo-Resistant Pancreatic Cancer","volume":"54","author":"Jayasinghe","year":"2022","journal-title":"Nat. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.cell.2012.12.012","article-title":"Intestinal Tumorigenesis Initiated by Dedifferentiation and Acquisition of Stem-Cell-like Properties","volume":"152","author":"Schwitalla","year":"2013","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1158\/0008-5472.CAN-06-2030","article-title":"Identification of Pancreatic Cancer Stem Cells","volume":"67","author":"Li","year":"2007","journal-title":"Cancer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e61","DOI":"10.1038\/onc.2012.28","article-title":"SOX2 Promotes Dedifferentiation and Imparts Stem Cell-like Features to Pancreatic Cancer Cells","volume":"2","author":"Zhang","year":"2013","journal-title":"Oncogenesis"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.canlet.2013.07.009","article-title":"Knockdown of Oct4 and Nanog Expression Inhibits the Stemness of Pancreatic Cancer Cells","volume":"340","author":"Lu","year":"2013","journal-title":"Cancer Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1038\/nrm1961","article-title":"The Biogenesis and Regulation of Telomerase Holoenzymes","volume":"7","author":"Collins","year":"2006","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.tibs.2013.07.001","article-title":"Telomerase: Central Regulator of All of the Hallmarks of Cancer","volume":"38","author":"Low","year":"2013","journal-title":"Trends Biochem. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1038\/nature08137","article-title":"Telomerase Modulates Wnt Signalling by Association with Target Gene Chromatin","volume":"460","author":"Park","year":"2009","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1270","DOI":"10.1038\/ncb2621","article-title":"Telomerase Directly Regulates NF-B-Dependent Transcription","volume":"14","author":"Ghosh","year":"2012","journal-title":"Nat. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1038\/s41586-020-1969-6","article-title":"Pan-Cancer Analysis of Whole Genomes","volume":"578","author":"Campbell","year":"2020","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1038\/ng.522","article-title":"A Genome-Wide Association Study Identifies Pancreatic Cancer Susceptibility Loci on Chromosomes 13q22.1, 1q32.1 and 5p15.33","volume":"42","author":"Petersen","year":"2010","journal-title":"Nat. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/s41586-021-03916-2","article-title":"Acinar Cell Clonal Expansion in Pancreas Homeostasis and Carcinogenesis","volume":"597","author":"Roake","year":"2021","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/j.cell.2018.03.034","article-title":"Machine Learning Identifies Stemness Features Associated with Oncogenic Dedifferentiation","volume":"173","author":"Malta","year":"2018","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.stemcr.2014.11.002","article-title":"A Flexible Reporter System for Direct Observation and Isolation of Cancer Stem Cells","volume":"4","author":"Tang","year":"2015","journal-title":"Stem Cell Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1038\/s41467-020-20474-9","article-title":"Integrated Analysis of Telomerase Enzymatic Activity Unravels an Association with Cancer Stemness and Proliferation","volume":"12","author":"Noureen","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Tang, R., Liu, X., Wang, W., Hua, J., Xu, J., Liang, C., Meng, Q., Liu, J., Zhang, B., and Yu, X. (2021). Identification of the Roles of a Stemness Index Based on MRNA Expression in the Prognosis and Metabolic Reprograming of Pancreatic Ductal Adenocarcinoma. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.643465"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1002\/path.5876","article-title":"Phosphorylation of HTERT at Threonine 249 Is a Novel Tumor Biomarker of Aggressive Cancer with Poor Prognosis in Multiple Organs","volume":"257","author":"Matsuda","year":"2022","journal-title":"J. Pathol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"6958","DOI":"10.1093\/emboj\/20.24.6958","article-title":"A Highly Selective Telomerase Inhibitor Limiting Human Cancer Cell Proliferation","volume":"20","author":"Damm","year":"2001","journal-title":"EMBO J."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Walter, K., Rodriguez-Aznar, E., Ventura Ferreira, M.S., Frappart, P.O., Dittrich, T., Tiwary, K., Meessen, S., Lerma, L., Daiss, N., and Schulte, L.A. (2021). Telomerase and Pluripotency Factors Jointly Regulate Stemness in Pancreatic Cancer Stem Cells. Cancers, 13.","DOI":"10.3390\/cancers13133145"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1764","DOI":"10.1038\/s41388-018-0553-0","article-title":"The Role of GLI-SOX2 Signaling Axis for Gemcitabine Resistance in Pancreatic Cancer","volume":"38","author":"Jia","year":"2019","journal-title":"Oncogene"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10586","DOI":"10.1038\/s41598-023-37836-0","article-title":"Telomere-Related Prognostic Biomarkers for Survival Assessments in Pancreatic Cancer","volume":"13","author":"Chen","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"36305","DOI":"10.18632\/oncotarget.16750","article-title":"Expression of Embryonal Stem Cell Transcription Factors in Breast Cancer: Oct4 as an Indicator for Poor Clinical Outcome and Tamoxifen Resistance","volume":"8","author":"Gwak","year":"2017","journal-title":"Oncotarget"},{"key":"ref_30","first-page":"bio034181","article-title":"Telomerase Reverse Transcriptase Coordinates with the Epithelial-to-Mesenchymal Transition through a Feedback Loop to Define Properties of Breast Cancer Stem Cells","volume":"7","author":"Ghoneim","year":"2018","journal-title":"Biol. Open"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2734","DOI":"10.1016\/j.cellsig.2013.08.041","article-title":"SOX2 Plays a Critical Role in EGFR-Mediated Self-Renewal of Human Prostate Cancer Stem-like Cells","volume":"25","author":"Rybak","year":"2013","journal-title":"Cell Signal"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2534","DOI":"10.1158\/0008-5472.CAN-16-1887","article-title":"WNT\/\u03b2-Catenin Directs Self-Renewal Symmetric Cell Division of HTERThigh Prostate Cancer Stem Cells","volume":"77","author":"Zhang","year":"2017","journal-title":"Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1410","DOI":"10.1038\/bjc.2011.94","article-title":"Downregulation of Transcription Factor SOX2 in Cancer Stem Cells Suppresses Growth and Metastasis of Lung Cancer","volume":"104","author":"Xiang","year":"2011","journal-title":"Br. J. Cancer"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1136\/jclinpath-2017-204739","article-title":"Cancer Stem Cells in Colorectal Cancer: A Review","volume":"71","author":"Munro","year":"2018","journal-title":"J. Clin. Pathol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2004320","DOI":"10.1002\/advs.202004320","article-title":"Colorectal Cancer Stem Cell States Uncovered by Simultaneous Single-Cell Analysis of Transcriptome and Telomeres","volume":"8","author":"Wang","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Bradshaw, A., Wickremsekera, A., Tan, S.T., Peng, L., Davis, P.F., and Itinteang, T. (2016). Cancer Stem Cell Hierarchy in Glioblastoma Multiforme. Front. Surg., 3.","DOI":"10.3389\/fsurg.2016.00021"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"P\u00e1dua, D., Barros, R., Amaral, A.L., Mesquita, P., Freire, A.F., Sousa, M., Maia, A.F., Caiado, I., Fernandes, H., and Pombinho, A. (2020). A SOX2 Reporter System Identifies Gastric Cancer Stem-like Cells Sensitive to Monensin. Cancers, 12.","DOI":"10.3390\/cancers12020495"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Vaddi, P., Stamnes, M., Cao, H., and Chen, S. (2019). Elimination of SOX2\/OCT4-Associated Prostate Cancer Stem Cells Blocks Tumor Development and Enhances Therapeutic Response. Cancers, 11.","DOI":"10.3390\/cancers11091331"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.cell.2011.11.025","article-title":"EMT and Dissemination Precede Pancreatic Tumor Formation","volume":"148","author":"Rhim","year":"2012","journal-title":"Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e08673","DOI":"10.1016\/j.heliyon.2021.e08673","article-title":"Human Telomerase Reverse Transcriptase Promotes the Epithelial to Mesenchymal Transition in Lung Cancer Cells by Enhancing C-MET Upregulation","volume":"8","author":"Prasad","year":"2022","journal-title":"Heliyon"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1016\/j.cell.2012.01.039","article-title":"Telomerase Reactivation Following Telomere Dysfunction Yields Murine Prostate Tumors with Bone Metastases","volume":"148","author":"Ding","year":"2012","journal-title":"Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"22886","DOI":"10.1038\/srep22886","article-title":"HTERT Promotes Cell Adhesion and Migration Independent of Telomerase Activity","volume":"6","author":"Liu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1038\/s41586-020-03147-x","article-title":"A Gene\u2013Environment-Induced Epigenetic Program Initiates Tumorigenesis","volume":"590","author":"Ho","year":"2021","journal-title":"Nature"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"eadd5327","DOI":"10.1126\/science.add5327","article-title":"Epigenetic Plasticity Cooperates with Cell-Cell Interactions to Direct Pancreatic Tumorigenesis","volume":"380","author":"Burdziak","year":"2023","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.cell.2019.03.010","article-title":"A Multiscale Map of the Stem Cell State in Pancreatic Adenocarcinoma","volume":"177","author":"Lytle","year":"2019","journal-title":"Cell"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.3892\/or.2015.3752","article-title":"Akt-Mediated Phosphorylation of Oct4 Is Associated with the Proliferation of Stem-like Cancer Cells","volume":"33","author":"Zhao","year":"2015","journal-title":"Oncol. Rep."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Rybak, A.P., Ingram, A.J., and Tang, D. (2013). Propagation of Human Prostate Cancer Stem-Like Cells Occurs through EGFR-Mediated ERK Activation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0061716"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"6632","DOI":"10.1158\/0008-5472.CAN-18-0650","article-title":"Circulating glioma cells exhibit stem cell-like properties Tianrun","volume":"78","author":"Liu","year":"2019","journal-title":"Cancer Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1002\/stem.2329","article-title":"Stem Cells Antigen-1 Enriches for a Cancer Stem Cell-Like Subpopulation in Mouse Gastric Cancer","volume":"34","author":"Park","year":"2016","journal-title":"Stem Cells"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Burchett, K.M., Yan, Y., and Ouellette, M.M. (2014). Telomerase Inhibitor Imetelstat (GRN163L) Limits the Lifespan of Human Pancreatic Cancer Cells. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0085155"},{"key":"ref_51","first-page":"657","article-title":"Telomere Shortening and Growth Inhibition of Human Cancer Cells by Novel Synthetic Telomerase Inhibitors MST-312, MST-295, and MST-1991","volume":"1","author":"Seimiya","year":"2002","journal-title":"Mol. Cancer Ther."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"3819","DOI":"10.1016\/j.febslet.2010.05.026","article-title":"Telomeres and Telomerase in Normal and Cancer Stem Cells","volume":"584","author":"Shay","year":"2010","journal-title":"FEBS Lett."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Rivero-Moreno, Y., Goyal, A., Bol\u00edvar, V., Osagwu, N., Echevarria, S., Gasca-Insuasti, J., Pereira-Graterol, F., von Ahrens, D., Gayt\u00e1n Fuentes, O.F., and Su\u00e1rez-Carre\u00f3n, L.O. (2025). Pancreaticobiliary Maljunction and Its Relationship with Biliary Cancer: An Updated and Comprehensive Systematic Review and Meta-Analysis on Behalf of TROGSS\u2014The Robotic Global Surgical Society. Cancers, 17.","DOI":"10.3390\/cancers17010122"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/2\/129\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T07:37:18Z","timestamp":1768203438000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/2\/129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,11]]},"references-count":53,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["cells15020129"],"URL":"https:\/\/doi.org\/10.3390\/cells15020129","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,11]]}}}