{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T17:12:59Z","timestamp":1790615579960,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,24]]},"group-title":"Cancer Biology","reference-count":78,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"funder":[{"award":["NIH R01 EY029795"],"award-info":[{"award-number":["NIH R01 EY029795"]}],"id":[{"id":"https:\/\/ror.org\/03wkg3b53","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Purpose<\/jats:title>\n                  <jats:p>Uveal melanoma (UM) is a rare but aggressive intraocular malignancy originating in the uvea (choroid, iris, and ciliary body). Despite primary treatment, 50% of UM patients develop metastases to the liver, lungs, bones, and skin. Topotecan, a topoisomerase I inhibitor, and SP141, an MDM2 inhibitor, have been studied in cancers, including retinoblastoma. This study explores a combined approach with these drugs in the 92-1 UM cell line.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    Primary choroidal melanoma-derived 92-1 cells were treated with topotecan and SP141 individually to determine IC\n                    <jats:sub>50<\/jats:sub>\n                    values. Cell viability and proliferation were assessed using the MTT assay and the xCELLigence real-time monitoring system. The synergistic effects were analyzed quantitatively using the Loewe Additivity and Highest Single-Agent (HSA) models. Flow cytometry was employed to analyze cell cycle distribution, while cell death and apoptosis were monitored via high-throughput, real-time quantitative analysis using the IncuCyte live-cell kinetic imaging system.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Both Topotecan and SP141 induced significant cell death in 92-1 cells. xCELLigence showed dose- and time-dependent inhibition of proliferation. Combining sub-IC\n                    <jats:sub>50<\/jats:sub>\n                    concentrations of topotecan and SP141 exhibited significant synergistic effects, markedly reducing growth. Cell cycle analysis revealed that topotecan causes a dose-dependent biphasic arrest in G2 and S phases, while SP141 causes G2\/M arrest, leading to cell death. Cytotoxicity assays corroborated increased cell death following combination therapy, and the Caspase 3\/7 assay indicated that apoptosis is the mechanism underlying cell death.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>The multi-targeted approach combining topotecan and SP141 drugs demonstrated synergistic therapeutic effects in 92-1 cells, suggesting a potential treatment strategy for managing UM.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.23.753860","type":"posted-content","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T11:25:15Z","timestamp":1790249115000},"source":"Crossref","is-referenced-by-count":0,"title":["Topotecan Synergizes with SP141 in the Management of Uveal Melanoma"],"prefix":"10.64898","author":[{"given":"Anju","family":"Thomas","sequence":"first","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Utkarsh R.","family":"Addi","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Audrianna","family":"Wu","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaleigh","family":"Kozak","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ross F.","family":"Collery","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amit","family":"Joshi","sequence":"additional","affiliation":[{"name":"Joint Department of Biomedical Engineering, Medical College of Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aparna","family":"Ramasubramanian","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"},{"name":"Children\u2019s Wisconsin, Milwaukee, WI 53226, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8725-676X","authenticated-orcid":false,"given":"Shyam S.","family":"Chaurasia","sequence":"additional","affiliation":[{"name":"Ocular Immunology & Angiogenesis Lab, Department of Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, WI 53226, USA"},{"name":"Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026092809200591000_2026.09.23.753860v1.1","doi-asserted-by":"publisher","DOI":"10.1002\/(sici)1097-0142"},{"key":"2026092809200591000_2026.09.23.753860v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/s0161-6420(98)92205-9"},{"key":"2026092809200591000_2026.09.23.753860v1.3","doi-asserted-by":"publisher","DOI":"10.1159\/000543151"},{"key":"2026092809200591000_2026.09.23.753860v1.4","unstructured":"Institute NC . 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