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男児外性器皮膚線維芽細胞におけるアンドロゲン標的候補遺伝子の探索

中尾 佳奈子 東北大学

2020.03.25

概要

テストステロンやジヒドロテストステロン(dihydrotestosterone:DHT)などの男性ホルモン(アンドロゲン)はアンドロゲン受容体(androgen receptor:AR)への結合を介して外性器男性化を促す。AR は核内受容体であり、リガンドとの結合によって細胞質から核内に移行する。そして標的遺伝子のアンドロゲン応答配列(androgen response element:ARE)に結合して転写制御を行う。これまでにヒト外性器におけるアンドロゲン標的遺伝子として APOD が同定されている。しかし、この遺伝子単独の作用だけでは複雑な外性器の男性化を完全には説明できず、詳細な外性器形成におけるアンドロゲン作用の分子メカニズムは未解明である。本研究は、ヒト外性器におけるアンドロゲン標的候補遺伝子を同定することを目的とした。

AR 変異 p.Ala597Thr を有する部分型アンドロゲン不応症患児 1 名および埋没陰茎男児 4 名(コントロール群)を対象とし、この 5 名の包皮由来皮膚線維芽細胞を用いてトランスクリプトーム解析を行った。まず各細胞を 2 つに分け、DHT またはメタノールを添加して培養した。その後、細胞から mRNAを抽出し、マイクロアレイ解析を行った。コントロール群において DHT 添加によって変動し、かつ、患者における変動がコントロール群よりも小さい遺伝子を抽出した。発現量が著しく低い遺伝子およびシグナルデータの品質が悪い遺伝子は解析対象から除外した。その結果、アンドロゲン標的候補遺伝子として 24 遺伝子が同定された。7 遺伝子が近傍に ARE 候補配列を有していた。24伝子の中には既知アンドロゲン標的遺伝子 APOD、マウス外性器原基男性化に関与すると報告されている CYP1B1、他臓器におけるアンドロゲン標的遺伝子として報告のある RGCC や HS6ST1 が含まれていた。コントロール群における 24 遺伝子の DHT 添加による log2 発現変動量は 0.3-0.7 であった。本研究の結果は、アンドロゲンによる外性器男性化の主体が、少数の特定遺伝子の大きな発現変動ではなく多数の遺伝子の微細変化であることを示唆する。

なお、本研究で同定された 24 個のアンドロゲン標的候補遺伝子には、これまでヒト疾患への関連が報告されていないものが多く含まれる。これらの遺伝子の変異は、臨床的にアンドロゲン不応症と診断されるが AR 変異が認められない患者における疾患原因となっている可能性がある。また本研究によって得られた知見は、AR 異常のためにアンドロゲン補充療法では十分な男性化が認められない患者のために、AR を介さずにアンドロゲン作用を発揮する新規治療薬の開発につながることが期待される。

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