講演情報
[O31-1]マウスを用いた経時的enhancer RNA解析による卵巣支持細胞enhancer候補領域の同定
○中川 竜一1, 高澤 啓1, 我有 茉希1, 辻 敦美2, 川路 英哉3,4, 村川 泰裕3, 高田 修治2, 三上 剛史5, 鳴海 覚志6, 深見 真紀6, 丸山 哲夫7, 林崎 良英8, 森尾 友宏1, 鹿島田 健一1 (1.東京医科歯科大学大学院 発生発達病態学分野, 2.国立成育医療研究センター システム発生・再生医学研究部, 3.理化学研究所 生命医科学研究センター, 4.東京都医学総合研究所 ゲノム医学研究センター, 5.国立成育医療研究センター データサイエンス部門 生物統計ユニット, 6.国立成育医療研究センター 分子内分泌研究部, 7.慶應義塾大学医学部 産婦人科, 8.理化学研究所 予防医療・診断技術開発プログラム)
In most cases of impaired ovarian development - such as 46,XX disorders of sex development (DSD) and premature ovarian insufficiency (POI) - the genetic causes have not been identified. It suggests that the majority of disease-associated variants could lie within non-coding sequences. In this study, we aimed to identify enhancers of ovarian genes known to play key roles in early ovarian development. We utilized bidirectional enhancer RNAs (eRNAs), of which transcriptional levels are shown to correlate with enhancer activities. The expression profiles in mouse WT1 positive fetal ovarian cells (=somatic cells) were obtained from cap analysis of gene expression (CAGE) at E13.5, E16.5 and P0. We compared the chronological expression profiles of ovarian specific intergenic RNA with expression profiles for each of the five ovarian genes, yielding two enhancer candidates (100 kb upstream of Wnt4 and 1 Mb upstream of Rspo1). Both sequences were well conserved between mouse and human, and we confirmed their enhancer activities using transient expression assays in murine granulosa cells. Further, by sequencing the region in patients (n=24 cases), such as POI, gonadal dysgenesis and 46,XX DSD, we identified rare single nucleotide variants (RSPO1 enhancer candidate: chr1:39249689C＞G, OR 27.15 (3.48-211.73), WNT4 enhancer candidate: chr1:22615333G＞A, OR 360.83 (21.9-5945)). Our results demonstrate that eRNA analysis presents a powerful tool for locating enhancers, and a means of identifying disease-associated variants, thus advancing an important unmet need in forward human genetics.