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卵巣癌患者におけるCirculating tumor DNAのDigital Droplet PCRを用いた測定は新たなバイオマーカーになりうる

湊 敬道 東北大学

2022.03.25

概要

卵巣癌は婦⼈科領域の難治性腫瘍であり,根治的⼿術や術後補助化学療法により腫瘍組織を寛解に⾄っても,数年以内に再発することが多い。従来の腫瘍マーカーであるCA125(Carbohydrate Antigen125)は、腫瘍切除後の経過観察に広く⽤いられているが、炎症などの他要因による影響を受けた場合に偽陽性を⽰し、また再発腫瘍が必ずしもCA125の上昇を伴わないため偽陰性の可能性もある。近年のliquid biopsyの発展により癌抗原マーカーのみならず、腫瘍細胞由来のDNA断⽚つまりCirculating tumor DNA(ctDNA)の検出が可能になった。ctDNAを計測することにより、卵巣癌再発の早期診断するための課題を克服することが期待されている。

本研究ではdigital droplet PCR(ddPCR)技術を⽤いて,寛解を得て経過観察中の卵巣癌患者のctDNAを計測した。対象となる患者には研究の説明を⾏い、同意を取得した。それぞれの患者からは、複数回にわたって連続的に外来受診の際、CA125の測定とctDNA検出⽤の⾎漿を採取した。対象患者の初回⼿術時に摘出した腫瘍組織から抽出したDNAの全エクソームシーケンスにより、腫瘍特異的な変異を同定し、患者ごとにddPCRプローブを選択した。

11例中6例が経過観察中に画像診断により再発を認め(平均無増悪⽣存期間は348.3⽇)、6例すべてにctDNAがddPCRにより検出された。その6例中5例でCA125の上昇前にctDNAをddPCRで検出した。ctDNAの対⽴遺伝⼦頻度の増加は、再発後の腫瘍量の増加と関連していた。また画像診断での再発⽇を基準として、CA125の増加よりも早くddPCRによりctDNAが検出された(ddPCRは49⽇前、CA125は7⽇後:p<0.05)。再発していない症例の⾎漿にはctDNAは検出されなかった。

本研究の結果は、ctDNAがCA125よりも鋭敏かつ定量的に卵巣癌の再発を反映することを⽰し、ctDNAが卵巣癌治療後のフォローアップにおいて理想的なマーカーであるという仮説を⽀持するものであった。

結論:ddPCRによるctDNAの測定は、卵巣癌再発の早期発⾒⽅法としてのバイオマーカーとなりうる。

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