tPA/tPA受容体シグナルによる多発性骨髄腫の病態制御機構の解明

• 研究課題/領域番号: 22KF0337
• 補助金の研究課題番号: 22F21773 (2022)
• 研究種目: 特別研究員奨励費
• 配分区分: 基金 (2023); 補助金 (2022)
• 応募区分: 外国
• 審査区分: 小区分54010:血液および腫瘍内科学関連
• 研究機関: 順天堂大学
• 研究代表者: ハイジッヒ ベアーテ 順天堂大学, 大学院医学研究科, 特任准教授 (30372931)
• 研究分担者: YATSENKO TETIANA 順天堂大学, 大学院医学研究科, 外国人特別研究員
• 研究期間 (年度): 2023-03-08 – 2024-03-31
• 研究課題ステータス: 交付 (2023年度)
• 配分額: 2,300千円 (直接経費: 2,300千円); 2023年度: 1,100千円 (直接経費: 1,100千円); 2022年度: 1,200千円 (直接経費: 1,200千円)
• キーワード: fibrinolysis / protein fragment / plasmin / multiple myeloma

研究開始時の研究の概要

Multiple myeloma (MM) requires neovascularization. Accumulating evidence suggests that the truncated tPA derivatives kringle 1-2 and kringle 2-protease domain have show antiangiogenic effect. Here, we will examine the role of kringle 2 of tPA during MM progression and it effects as an anti-angiogenic drug. In addition, we will test its anti-proliferative effects on myeloma cell growth and establish its effects on the coagulation system through its potential interactions with platelets.

研究実績の概要

This study aimed to find an efficient method to generate tissue-type plasminogen activator (tPA) fragments, which naturally occur during tumor progression/inflammation of the hematologic cancer called multiple myeloma. To determine whether it will occur. In the first year of this proposal, we used different approaches to generate tPA fragments. Using a plasmid-based expression system in bacteria, we evaluated multiple proteases, including alpha elastase and plasmin, to cleave the purchased protein. We have successfully expressed the fragment in bacteria and are currently evaluating various methods for large-scale isolation. In one of his original articles published this year, we identified p53 as a downstream target of the small molecule and plasmin inhibitor YO2 (Cancers 15, 288; doi 10.3390/cancers15010288, 2023). A published review article summarizes the knowledge of the plasminogen-plasmin system in the oral cavity (Cells, 12(3), 445 https://doi.org/10.3390/cells12030445, 2023)."

現在までの達成度 (区分)

2: おおむね順調に進展している

理由

We have initially used recombinant tPA and performed time course studies to identify proteases that will generate the tPA fragments. Unfortunately, non of the tested proteases was able to generate sufficient hydrolysis of the input full-length protein. Therefore, we shifted the strategy to express the tPA fragment in a plasmid. The generation of the fragment in bacteria turned out to be more difficult than expect, but we identified a strain in which the protein could be expressed. We are currently using various purification methods to obtain the protein in a tissue culture pure form.

今後の研究の推進方策

We are planning in the following year to continue our studies on the role of the tPA fragment in tumor progression* 1. Purification of the tPA fragment
We already have expressed the protein in an expression vector and obtained supernatants containing the protein fragments. 2. Identification of the natural occurrence of the tPA fragment(s) in selected hematological and solid cancers using Western blotting in cell lines in vitro and murine disease models in vivo. 3. Identification of fragment properties during cancer progression with focus on cell proliferation and migration. A solid cancer and the hematological disease multiple myeloma will be targeted. An orthotopic murine model for the solid cancer has been established in the lab. Furthermore, the multiple myeloma model as described in Salama et al Blood advances 2020) will be used in the current study. 4. Evaluation of the anti-angiogenic properties of the newly synthesized fragment。

研究成果

• [国際共同研究] University of Cologn/University Hospital of Frankfurt/University Hospital of Augsburg(ドイツ)
• [国際共同研究] Federal University of Bahia(ブラジル)
• [国際共同研究] An-Najah National University(パレスチナ(PLO))
• [雑誌論文] The Role of the Plasminogen/Plasmin System in Inflammation of the Oral Cavity (2023)
  • 著者名/発表者名: Yatsenko Tetiana、Skrypnyk Maksym、Troyanovska Olga、Tobita Morikuni、Osada Taro、Takahashi Satoshi、Hattori Koichi、Heissig Beate
  • 雑誌名: Cells 巻: 12 号: 3 ページ: 445-445
  • DOI: 10.3390/cells12030445
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] YO2 Induces Melanoma Cell Apoptosis through p53-Mediated LRP1 Downregulation (2022)
  • 著者名/発表者名: Salama Yousef、Takahashi Satoshi、Tsuda Yuko、Okada Yoshio、Hattori Koichi、Heissig Beate
  • 雑誌名: Cancers 巻: 15 号: 1 ページ: 288-288
  • DOI: 10.3390/cancers15010288
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] COVID-19 severity and thrombo-inflammatory response linked to ethnicity (2022)
  • 著者名/発表者名: Heissig B, Salama Y, Vehreschild JJ, Vehreschild MJGT, Mori H, Adachi E, Jakob C, Tabe Y, Ruethrich M, Borgmann S, Naito T, Wille K, Valenti S, Hower M, Hattori N, Rieg S, Nagaoka T, Jensen BE, Yotsuyanagi H, Hertenstein B, Ogawa H, Kominami E, Roemmele C, Takahashi S, Rupp J, Takahashi K, Hanses F, Hattori K
  • 雑誌名: Biomedicines 巻: 10 号: 10 ページ: 2549-2549
  • DOI: 10.3390/biomedicines10102549
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] siRNA against CD40 delivered via a fungal recognition receptor ameliorates murine acute graft-versus-host disease (2022)
  • 著者名/発表者名: Heissig B, Salama Y, Tateno M, Takahashi S and Hattori K
  • 雑誌名: EJHaem 巻: 3 号: 3 ページ: 849-861
  • DOI: 10.1002/jha2.439
  • 査読あり / オープンアクセス / 国際共著
• [雑誌論文] アンジオクラインシステムによるがん増殖制御機構 (2022)
  • 著者名/発表者名: 服部浩一、島津浩、高橋聡、Heissig Beate
  • 雑誌名: 日本血栓止血学会雑誌 巻: 33 ページ: 314-320
• [雑誌論文] サイトカインストーム関連疾患に対する新規分子標的としての血液線維素溶解系因子群 (2022)
  • 著者名/発表者名: 服部浩一、島津浩、高橋聡、Heissig Beate
  • 雑誌名: 臨床血液 巻: 63 ページ: 403-409
• [学会発表] COVID-19後のプロテオリティック・ニッチにおける免疫系再構築機構の解明 (2023)
  • 著者名/発表者名: 服部浩一、高橋聡、長田太郎、Heissig Beate
  • 学会等名: 第22回日本再生医療学会総会
• [学会発表] ES特異的転写因子Zfp296の細胞がん化への関与 (2022)
  • 著者名/発表者名: 溝上優美、池田智美、池上貴子、服部浩一、Heissig Beate 小出寛
  • 学会等名: 第95回日本生化学会大会
• [学会発表] Biomarker and predictors of severe COVID-19:a comparative study of patients in Japan and Germany (2022)
  • 著者名/発表者名: Hattori K,Takahashi S,and Heissig Beate
  • 学会等名: 第84回日本血液学会学術集会
• [学会発表] サイトカインストーム症候群の病態解明と新規治療戦略 (2022)
  • 著者名/発表者名: 服部浩一、高橋聡、Heissig Beate
  • 学会等名: 第44回日本造血・免疫細胞療法学会