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We analyzed RNA expression profiles of 15 epigenetic mediators in uterine fibroids compared to myometrium using publicly available RNA sequencing (RNA-seq) data. To validate our findings, we performed RT-qPCR on a separate cohort of uterine fibroids targeting these modifiers confirming our RNA-seq data. We then examined protein profiles of key N6-methyladenosine (m6A) modifiers in fibroids and their matched myometrium, showing no significant differences in concordance with our RNA expression profiles. To determine RNA modification abundance, mRNA and small RNA from fibroids and matched myometrium were analyzed by ultra-high performance liquid chromatography-mass spectrometry identifying prevalent m6A and 11 other known modifiers. However, no aberrant expression in fibroids was detected. We then mined a previously published dataset and identified differential expression of m6A modifiers that were specific to fibroid genetic subtype. Our analysis also identified m6A consensus motifs on genes previously identified to be dysregulated in uterine fibroids. Overall, using state-of-the-art mass spectrometry, RNA expression, and protein profiles, we characterized and identified differentially expressed m6A modifiers in relation to driver mutations. Despite the use of several different approaches, we identified limited differential expression of RNA modifiers and associated modifications in uterine fibroids. However, considering the highly heterogenous genomic and cellular nature of fibroids, and the possible contribution of single molecule m6A modifications to fibroid pathology, there is a need for greater in-depth characterization of m6A marks and modifiers in a larger and diverse patient cohort.<\/jats:p>","DOI":"10.1210\/endocr\/bqae074","type":"journal-article","created":{"date-parts":[[2024,6,28]],"date-time":"2024-06-28T00:31:37Z","timestamp":1719534697000},"source":"Crossref","is-referenced-by-count":4,"title":["Characterization of m6A Modifiers and RNA Modifications in Uterine Fibroids"],"prefix":"10.1210","volume":"165","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0943-4422","authenticated-orcid":false,"given":"Jitu W","family":"George","sequence":"first","affiliation":[{"name":"Olson Center for Women's Health, Department of Obstetrics and Gynecology, University of Nebraska Medical Center , Omaha, NE 68198 , USA"},{"name":"Veterans Affairs Nebraska Western Iowa Health Care System , Omaha, NE 68105 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosa A","family":"Cancino","sequence":"additional","affiliation":[{"name":"Olson Center for Women's Health, Department of Obstetrics and Gynecology, University of Nebraska Medical Center , Omaha, NE 68198 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jennifer L","family":"Griffin Miller","sequence":"additional","affiliation":[{"name":"Olson Center for Women's Health, Department of Obstetrics and Gynecology, University of Nebraska Medical Center , Omaha, NE 68198 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fang","family":"Qiu","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, University of Nebraska Medical Center , Omaha, NE 68198 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qishan","family":"Lin","sequence":"additional","affiliation":[{"name":"RNA Epitranscriptomics and Proteomics Resource, Department of Chemistry, University at Albany , Albany, NY 12222 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M Jordan","family":"Rowley","sequence":"additional","affiliation":[{"name":"Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center , Omaha, NE 68198 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Varghese M","family":"Chennathukuzhi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Integrative Physiology, University of Kansas Medical Center , Kansas City, KS 66160 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3468-4079","authenticated-orcid":false,"given":"John S","family":"Davis","sequence":"additional","affiliation":[{"name":"Olson Center for Women's Health, Department of Obstetrics and Gynecology, University of Nebraska Medical Center , Omaha, NE 68198 , USA"},{"name":"Veterans Affairs Nebraska Western Iowa Health Care System , Omaha, NE 68105 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"80","published-online":{"date-parts":[[2024,7,1]]},"reference":[{"issue":"2","key":"2024072604374596900_bqae074-B1","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1055\/s-0037-1599090","article-title":"Recent advances in uterine fibroid etiology","volume":"35","author":"McWilliams","year":"2017","journal-title":"Semin Reprod Med"},{"issue":"5","key":"2024072604374596900_bqae074-B2","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1093\/humupd\/dmv030","article-title":"Epidemiological and genetic clues for molecular mechanisms involved in uterine leiomyoma development and growth","volume":"21","author":"Commandeur","year":"2015","journal-title":"Hum Reprod Update"},{"issue":"4","key":"2024072604374596900_bqae074-B3","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1093\/ajcp\/94.4.435","article-title":"The frequency of uterine 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