{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T00:32:00Z","timestamp":1792801920320,"version":"4.5.0"},"posted":{"date-parts":[[2026,5,24]]},"group-title":"Cancer Biology","reference-count":51,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,24]],"date-time":"2026-05-24T00:00:00Z","timestamp":1779580800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100017288","name":"Bradens\u2019 Hope For Childhood Cancer","doi-asserted-by":"crossref","award":["Pilot Grant"],"award-info":[{"award-number":["Pilot Grant"]}],"id":[{"id":"10.13039\/100017288","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Massman\u2019s family research Fund","award":["40374100"],"award-info":[{"award-number":["40374100"]}]},{"DOI":"10.13039\/100012640","name":"Alan B Slifka Foundation","doi-asserted-by":"crossref","award":["Research Fund"],"award-info":[{"award-number":["Research Fund"]}],"id":[{"id":"10.13039\/100012640","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Cancer Institute","award":["P30 CA168524"],"award-info":[{"award-number":["P30 CA168524"]}]},{"award":["General Research Fund (GRF)"],"award-info":[{"award-number":["General Research Fund (GRF)"]}],"id":[{"id":"https:\/\/ror.org\/001tmjg57","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Ewing sarcoma is a pediatric cancer that develops in skeletal elements. The majority of Ewing sarcoma patients carry the aberrant\n                  <jats:italic>EWSR1-FLI1<\/jats:italic>\n                  fusion gene. Despite trisomy 8 being an additional common aberration associated with a poor prognosis for patients, its induction mechanism remains unknown. When the\n                  <jats:italic>EWSR1-FLI1<\/jats:italic>\n                  gene is formed, the cell loses one wildtype EWSR1 allele. To elucidate the induction mechanism of trisomy 8, we generated a cell line that allows for the conditional induction of EWSR1-FLI1 expression and EWSR1 knockdown (derived from a single\n                  <jats:italic>EWSR1<\/jats:italic>\n                  allele. Specifically, the conditional cell line was generated by integrating the Tet-on\n                  <jats:italic>EWSR1-FLI1<\/jats:italic>\n                  construct into the\n                  <jats:italic>AAVS<\/jats:italic>\n                  locus and adding a\n                  <jats:italic>miniAID<\/jats:italic>\n                  tag at the 5\u2019 end of the EWSR1 locus using auxin-degron system. A combination of the EWSR1-FLI1 expression and degradation of one allele-derived EWSR1 induced a high incidence of trisomy 8 within eight days, enhancing colony formation. Mechanistically, trisomy 8 is induced by the haploinsufficiency of EWSR1, and the remaining EWSR1 proteins are likely inhibited by interaction with EWSR1-FLI1. Our data showed that the knockout of EWSR1 alone was sufficient to increase the incidence of trisomy 8. Expression of wild-type EWSR1 in EWSR1 knockout cells rescued the high incidence of trisomy 8. In contrast, the EWSR1:R565A mutant, which lacks the ability to interact with Aurora B kinase, failed to rescue this phenotype. We propose that the combination of EWSR1-FLI1 expression and loss of EWSR1 contributes to the induction of trisomy 8 through the compromised EWSR1-Aurora B pathway.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Graphical Abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" fig-type=\"figure\" orientation=\"portrait\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"726567v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.21.726567","type":"posted-content","created":{"date-parts":[[2026,5,24]],"date-time":"2026-05-24T23:30:13Z","timestamp":1779665413000},"source":"Crossref","is-referenced-by-count":2,"title":["The combination of\n                  <i>EWSR1-FLI1<\/i>\n                  and loss of one\n                  <i>EWSR1<\/i>\n                  allele leads to the induction of trisomy 8"],"prefix":"10.64898","author":[{"given":"Harsha","family":"Hapugaswatta","sequence":"first","affiliation":[{"name":"University of Kansas, Molecular Biosciences, Lawrence KS, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Parrales","sequence":"additional","affiliation":[{"name":"Children\u2019s Mercy Research Institute, Department of Pediatrics, Kansas City MO, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyewon","family":"Park","sequence":"additional","affiliation":[{"name":"University of Kansas, Molecular Biosciences, Lawrence KS, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haeyoung","family":"Kim","sequence":"additional","affiliation":[{"name":"University of Kansas, Molecular Biosciences, Lawrence KS, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomoo","family":"Iwakuma","sequence":"additional","affiliation":[{"name":"Children\u2019s Mercy Research Institute, Department of Pediatrics, Kansas City MO, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mizuki","family":"Azuma","sequence":"additional","affiliation":[{"name":"University of Kansas, Molecular Biosciences, Lawrence KS, Unites States"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052705052374000_2026.05.21.726567v1.1","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.3389\/fonc.2019.01043","article-title":"Risk factors for metastasis at initial diagnosis with Ewing sarcoma","volume":"9","year":"2019","journal-title":"Frontiers in Oncology"},{"key":"2026052705052374000_2026.05.21.726567v1.2","doi-asserted-by":"publisher","DOI":"10.1002\/pbc.21618"},{"key":"2026052705052374000_2026.05.21.726567v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/359162a0"},{"key":"2026052705052374000_2026.05.21.726567v1.4","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199408043310503"},{"key":"2026052705052374000_2026.05.21.726567v1.5","first-page":"1229","article-title":"A variant Ewing\u2019s sarcoma translocation (7;22) fuses the EWS gene to the ETS gene ETV1","volume":"10","year":"1995","journal-title":"Oncogene"},{"key":"2026052705052374000_2026.05.21.726567v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1200933"},{"key":"2026052705052374000_2026.05.21.726567v1.7","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-2264(199602)15:2<115::AID-GCC6>3.0.CO;2-6"},{"key":"2026052705052374000_2026.05.21.726567v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2016.03.009"},{"key":"2026052705052374000_2026.05.21.726567v1.9","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/362173"},{"key":"2026052705052374000_2026.05.21.726567v1.10","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-013-9427-7"},{"key":"2026052705052374000_2026.05.21.726567v1.11","doi-asserted-by":"crossref","unstructured":"Y. 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