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Characterization of m6A Modifiers and RNA Modifications in Uterine Fibroids

子宫肌瘤 肌层 核糖核酸 生物 基因表达 平滑肌瘤 RNA序列 转录组 计算生物学 基因 病理 内分泌学 子宫 遗传学 医学
作者
Jitu W. George,Rosa A. Cancino,Jennifer L. Griffin Miller,Fang Qiu,Qishan Lin,M. Jordan Rowley,Varghese M. Chennathukuzhi,John S. Davis
出处
期刊:Endocrinology [The Endocrine Society]
卷期号:165 (8) 被引量:3
标识
DOI:10.1210/endocr/bqae074
摘要

Abstract Uterine leiomyoma or fibroids are prevalent noncancerous tumors of the uterine muscle layer, yet their origin and development remain poorly understood. 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.
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