Developments of HE, fluorescent immunostaining, and WSI analysis by FISH and applications to Hodgkin lymphoma

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K08746
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Human pathology
• Research Institution: Nagoya City University
• Principal Investigator: Murase Takayuki 名古屋市立大学, 医薬学総合研究院(医学), 准教授 (40315875)
• Co-Investigator(Kenkyū-buntansha): 稲垣 宏 名古屋市立大学, 医薬学総合研究院(医学), 教授 (30232507)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 蛍光 in situ hybridization / Sequential FICTION-WSI / 免疫染色 / IN-Cell Analyzer / 顕微鏡画像解析 / 染色体転座 / 多形腺腫 / 多発性骨髄腫

Outline of Final Research Achievements

The CCND1-IGH, NSD2-IGH, IGH-MAF gene rearrangements by FISH are used for risk stratification in multiple myeloma (MM) cases. Compared with fresh cell-FISH, immunohistochemistry (IHC) is more cost-/time-efficient and can be readily applied to routinely prepared FFPE materials. We performed FFPE-FISH and FFPE-IHC, and examined the usefulness of IHC as a tool for detecting CCND1, NSD2, and MAF gene rearrangements. With cohort n=70, we performed FFPE-FISH and FFPE-IHC, and determined IHC cut-off points. The sensitivity and specificity for the molecules were >.90 and >.96, respectively. With cohort n=120 using unknown gene status cases, we performed IHC, and the gene status was estimated using the cut-off points determined with cohort n=70. The subsequent FISH analysis showed that the sensitivity and specificity for the molecules were >.92 and >.98, respectively. The MM gene rearrangements were estimated accurately by IHC, suggesting that fresh cell-FISH assays can be replaced by IHC.

Free Research Field

分子病理診断学

Academic Significance and Societal Importance of the Research Achievements

次世代シーケンサーにより網羅的に遺伝子を解析し、腫瘍遺伝子異常をビッグデータ化することは腫瘍発生研究に重要であるが、腫瘍組織のビッグデータとの比較の報告はない。我々は1枚のパラフィン組織切片から蛍光免疫染色とFISH法を連続的に行い、スライド全体をビッグデータ化し、細胞単位で組織全体を解析する技術(Sequential FICTION-WSI法)を開発したが、基礎医学・臨床医学分野での応用が可能である。以上から遺伝子ビッグデータと細胞単位の形態・蛋白発現・遺伝子解析から得られた組織ビッグデータとを比較することは画期的であり、腫瘍組織を対象とした分子病理学の応用範囲を飛躍的に広げるものである。