血中循環腫瘍細胞を標的とした新規肝がんリスク診断法の開発

2022 Fiscal Year Research-status Report

• Project/Area Number: 21K15944
• Research Institution: University of Fukui
• Principal Investigator: 大藤 和也 福井大学, 学術研究院医学系部門, 特別研究員 (60597699)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 肝細胞癌 / 血中循環腫瘍細胞

Outline of Annual Research Achievements

本研究では肝細胞がん患者における薬物療法前後での循環腫瘍細胞(CTC)の定量性、及びがん進展関連遺伝子の発現を網羅的に解析することで肝細胞がんの進展・薬物療法による早期治療効果の判定に関わる新規バイオマーカーの同定や評価法開発を目的とした。
Atezolizumab-Bevacizumab併用療法肝がん症例の治療前，3週後，効果判定時に末梢血よりRosetteSep（STEMCELL Technologies）でCTCを濃縮，BD FACS Aria IIを用いて細胞表面マーカー（CD45，CD90，CD133，PanCK，EpCAM，Vimentin）を解析，CD45陰性かつPanCK陽性細胞をCTCと定義した．CTCからRNAを抽出しがん進展に関連する373遺伝子の発現変化を次世代シーケンサー（NGS）で解析，GSEA（Gene Set Enrichment Analysis）でパスウェイ解析を行った．
PR群のCTC数は治療経過で減少し（p=0.01），効果判定時はPD群と比較し有意に低値であった（p＜0.01）．細胞サブセット解析でPD群は治療前Vimentin陽性CTC数が多かった．NGS解析ではPR群のCTCはアポトーシス経路関連遺伝子（FAS，TP53）の発現が，PD群ではTGF-β経路関連遺伝子（TGF-β1，SMAD2）の発現が有意に上昇していた（p＜0.05）．
肝がん複合免疫療法において治療効果に応じてCTC数の変動を認めた．さらにCTC発現解析により細胞内アポトーシス経路やTGF-β経路に関連する遺伝子発現が治療効果と相関することが示された．以上の結果より肝がん複合免疫療法中における循環腫瘍細胞はがん進展・治療耐性を予見する有用なバイオマーカーとなる可能性が示唆された．

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

CTC解析について比較的順調に症例蓄積がなされている。

Strategy for Future Research Activity

症例をさらに蓄積するとともに、肝がん進展に関わる因子としてCTCにおける上皮間葉転換，幹細胞能や増殖能に関連する遺伝子発現等についても網羅的遺伝子発現解析を検討している。

Research Products

• [Journal Article] Histology and cytokine levels in hepatic injury accompanying a case of non-severe COVID-19. (2023)
  • Author(s): Matsuda H, Nosaka T, Hiramatsu K, Takahashi K, Naito T, Ofuji K, Ohtani M, Imamura Y, Iwasaki H, Nakamoto Y.
  • Journal Title: Clin J Gastroenterol. Volume: 16 Pages: 270-278
  • DOI: 10.1007/s12328-023-01755-4.
  • Peer Reviewed / Open Access
• [Journal Article] Hepatocellular carcinoma progression promoted by 5-lipoxygenase activity in CD163(+) tumor-associated macrophages. (2023)
  • Author(s): Nosaka T, Murata Y, Takahashi K, Naito T, Ofuji K, Matsuda H, Ohtani M, Hiramatsu K, Imamura Y, Goi T, Nakamoto Y.
  • Journal Title: Biomed Pharmacother. Volume: 162 Pages: 114592
  • DOI: 10.1016/j.biopha.2023.114592.
  • Peer Reviewed / Open Access
• [Journal Article] Elderly onset congenital hepatic fibrosis with portal hypertension diagnosed after recurrent cholangitis: a case report. (2022)
  • Author(s): Takahashi K, Ofuji K, Nosaka T, Naito T, Matsuda H, Ohtani M, Hiramatsu K, Harada K, Naiki H, Nakamoto Y.
  • Journal Title: Clin J Gastroenterol. Volume: 15(3) Pages: 611-616
  • DOI: 10.1007/s12328-022-01620-w.
• [Journal Article] Successful early diagnosis of monomorphic epitheliotropic intestinal T-cell lymphoma manifesting as chronic diarrhea and hypokalemia using video capsule endoscopy and double-balloon enteroscopy. (2022)
  • Author(s): Naito T, Nosaka T, Takahashi K, Ofuji K, Matsuda H, Ohtani M, Hiramatsu K, Imamura Y, Yamauchi T, Nakamoto Y.
  • Journal Title: Clin J Gastroenterol. Volume: 15(5) Pages: 913-919
  • DOI: 10.1007/s12328-022-01676-8.
• [Journal Article] A rare case of eosinophilic gastritis induced by nivolumab therapy for metastatic melanoma. (2022)
  • Author(s): Tsuji A, Hiramatsu K, Namikawa S, Yamamoto A, Midori Y, Murata Y, Tanaka T, Nosaka T, Naito T, Takahashi K, Ofuji K, Matsuda H, Ohtani M, Imamura Y, Iino S, Hasegawa M, Nakamoto Y.
  • Journal Title: Clin J Gastroenterol. Volume: 15(5) Pages: 876-880
  • DOI: 10.1007/s12328-022-01680-y.
• [Journal Article] Cholesterol crystal embolism in multiple organs after transarterial chemoembolization for hepatocellular carcinoma: An autopsy case report. (2022)
  • Author(s): Yamashita J, Nosaka T, Takahashi K, Naito T, Ofuji K, Matsuda H, Ohtani M, Hiramatsu K, Kobayashi M, Nakamoto Y.
  • Journal Title: Medicine (Baltimore) Volume: 101(39) Pages: e30769
  • DOI: 10.1097/MD.0000000000030769.
• [Presentation] Clinical Usefulness of Skeletal Muscle Quantity as Prognostic Factor in Elderly Patients with Advanced Pancreatic Cancer Receiving Nab-Paclitaxel Plus Gemcitabine Chemotherapy (2022)
  • Author(s): Akazawa Y, Ohtani M, Nosaka T, Takahashi K, Naito T, Ofuji K, Matsuda H, Hiramatsu K, Nakamoto Y
  • Organizer: DDW 2022
  • Int'l Joint Research
• [Presentation] Risk Factors for Pancreatic Malignancies in Patients with Low-Risk Branch Duct Intraductal Papillary Mucinous Neoplasms after 5 Years of Long-Term Follow-Up (2022)
  • Author(s): Ohtani M, Ofuji K, Akazawa Y, Nosaka T, Takahashi K, Naito T, Matsuda H, Hiramatsu K, Nakamoto Y
  • Organizer: DDW 2022
  • Int'l Joint Research
• [Presentation] Increased Liver Volume Measured using a 3d Medical Imaging in Patients with Advanced Decompensated Cirrhosis Treated with Balloon-Occluded Retrograde Transvenous Obliteration (B-Rto) (2022)
  • Author(s): Yamamoto A, Ofuji K, Nosaka T, Akazawa Y, Naito T, Takahashi K, Matsuda H, Ohtani M, Nakamoto Y
  • Organizer: AASLD 2022
  • Int'l Joint Research
• [Presentation] Stem Cell Signature of Peripheral Circulating Tumor Cells correlated with therapeutic effects of Atezolizumab plus Bevacizumab in unresectable HCC patients (2022)
  • Author(s): Murata Y, Nosaka T, Takahashi K, Naito T, Ofuji K, Matsuda H, Ohtani M, Nakamoto Y
  • Organizer: AASLD 2022
  • Int'l Joint Research