{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T23:56:03Z","timestamp":1790812563025,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,27]]},"group-title":"Molecular Biology","reference-count":33,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T00:00:00Z","timestamp":1790467200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","id":[{"id":"https:\/\/ror.org\/04byxyr05","id-type":"ROR","asserted-by":"crossref"}]},{"name":"ASCI","award":["PSSF Fellowship"],"award-info":[{"award-number":["PSSF Fellowship"]}]},{"name":"UC Davis","award":["ARC-MD"],"award-info":[{"award-number":["ARC-MD"]}],"id":[{"id":"https:\/\/ror.org\/05rrcem69","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Endometriosis is an estrogen-driven, inflammatory disorder affecting \u223c10% of menstruators, causing severe pain. Current treatments that reduce estrogen or inflammation have inconsistent efficacy and poor tolerability. Therapies identified via drug-repositioning methods, which target patient-specific pathways, offer a promising alternative. Simvastatin and primaquine were identified as potential treatments, strongly reversing endometriosis-associated gene expression pathways, and behavioral testing in an animal model showed that these drugs diminish endometriosis-associated pain. This paper examines their effects in human assembloids to provide proof-of-concept validation and insights into their mechanisms of action relevant to endometriosis.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We established human endometrial assembloids using immortalized epithelial (12Z) and stromal fibroblast (iEc) cell lines, as well as primary tissue from patients. Assembloids were generated in 96-well agarose molds with both cell types. We tested ibuprofen, fenoprofen, primaquine, and simvastatin. Following a 24-hour exposure, assembloids and monolayer cultures were harvested and stranded mRNA-seq libraries were sequenced on an Illumina NovaSeqX Plus System.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Gene set enrichment analysis showed key pathway reversals: Primaquine reversed the chemical carcinogenesis ROS pathway in assembloids. Simvastatin reversed cytokine-cytokine receptor interaction in both epithelial and stromal cell lines, and the cytoskeleton in muscle cells pathway in stromal cell lines. Fenoprofen reversed the calcium signaling pathway in stromal cell lines.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>Simvastatin induced the most significant gene expression changes, notably reversing cytokine\u2013cytokine receptor interactions in both cell lines, mirroring findings from our rat model. Despite the small sample size limitation, these experiments highlight the promise of assembloid models to test therapeutic candidates for endometriosis.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.27.754752","type":"posted-content","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T04:40:14Z","timestamp":1790570414000},"source":"Crossref","is-referenced-by-count":0,"title":["Simvastatin and Primaquine Identified as Potential Endometriosis Therapeutics via a Novel Epithelial-Stromal Assembloid Drug Screening Assay"],"prefix":"10.64898","author":[{"given":"Ferheen","family":"Abbasi","sequence":"first","affiliation":[{"name":"Department of Obstetrics, Gynecology & Reproductive Sciences, UCSF, San Francisco, CA, USA"},{"name":"School of Medicine, UC Davis, Sacramento, CA, USA"},{"name":"Mass General Brigham Hospital, Harvard University, Boston, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4567-4012","authenticated-orcid":false,"given":"Xinyu","family":"Tang","sequence":"additional","affiliation":[{"name":"Bakar Computational Health Sciences Institute, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan C.","family":"Irwin","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynecology & Reproductive Sciences, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Binya","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynecology & Reproductive Sciences, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7393-5120","authenticated-orcid":false,"given":"Tomiko T.","family":"Oskotsky","sequence":"additional","affiliation":[{"name":"Bakar Computational Health Sciences Institute, UCSF, San Francisco, CA, USA"},{"name":"Division of Clinical Informatics and Digital Transformation, Department of Medicine, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7246-6083","authenticated-orcid":false,"given":"Marina","family":"Sirota","sequence":"additional","affiliation":[{"name":"Bakar Computational Health Sciences Institute, UCSF, San Francisco, CA, USA"},{"name":"Department of Pediatrics, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederick J.","family":"Meyers","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Hematology and Oncology, UC Davis Health, Sacramento, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1677-0822","authenticated-orcid":false,"given":"Linda C.","family":"Giudice","sequence":"additional","affiliation":[{"name":"Department of Obstetrics, Gynecology & Reproductive Sciences, UCSF, San Francisco, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026093016151148000_2026.09.27.754752v1.1","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1097\/GRF.0000000000000292","article-title":"Medical Management of Endometriosis","volume":"60","year":"2017","journal-title":"Clinical Obstetrics & Gynecology"},{"key":"2026093016151148000_2026.09.27.754752v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/J.FERTNSTERT.2017.05.004"},{"key":"2026093016151148000_2026.09.27.754752v1.3","doi-asserted-by":"crossref","first-page":"2946","DOI":"10.1007\/s43032-025-01947-y","article-title":"Factors Affecting Hormonal Therapy Ineffectiveness and Discontinuation due to Adverse Effects in Patients with Endometriosis-Associated Pain","volume":"32","year":"2025","journal-title":"Reprod Sci"},{"key":"2026093016151148000_2026.09.27.754752v1.4","doi-asserted-by":"crossref","unstructured":"Chen Y , Waseem S , Luo L . 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