Therapeutic potential of novel antibody-oligonucleotide conjugate for intractable B-cell lymphomas

新型抗体-寡核苷酸缀合物治疗难治性 B 细胞淋巴瘤的潜力

• 批准号: 22K08497
• 负责人: 長尾 俊景
• 金额: $ 2.58万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22K08497/
• 关键词: 難治性B細胞腫瘍; 細胞内浸透抗体; S化CpG-ODN; 抗体-オリゴヌクレオチド複合体

当該年度（令和4年度）の研究内容の当初の目標としては、本研究のコンセプトである、抗体-S化CpG-ODN複合体の実験系の検証と確立であった。すなわち、最終的な目標はB細胞リンパ腫の病態特異的細胞内蛋白に対する抗体に、S化CpG-ODN複合体をconjugateし、細胞内への浸透性を付与した抗体-オリゴヌクレオチド複合体（AOC）を作成し、その抗腫瘍効果（細胞内TLR9を通じた）を検証することであるが、まずは同様の構造を持ち、比較的描出が容易な細胞内蛋白（細胞骨格蛋白など）を標的としたモデルAOCを作成し、その細胞内分布や局在性を確認することであった。実績：代表的な細胞骨格蛋白であるβ-actin（あるいはα-tubulin）に対するマウスモノクローナル抗体に、商業ベースでオーダー可能な蛍光色素標識Ｓ化ODNを、アフィニティー法（ストレプトアビジン-ビオチン）を用いてconjugateした後、当該AOCを純化し、試験的なモデル試薬とした。品質や濃度の安定が困難であったが、最終的に様々な細胞株（上皮系悪性腫瘍、造血器腫瘍、293T細胞等）への投与・インキュベートを試み、十分な洗浄の後フローサイトメトリー法を用いて蛍光色素の細胞内分布について評価した。しかし結果的に陽性コントロールと比較して蛍光色素の細胞内信号強度は明らかに低く、一方蛍光色素標識S化ODNのみによる細胞処理の場合（非特異的反応）と比較しても、有意差を見出せなかった。これらの結果が検証実験の方法や、試薬の作成過程における技術面での複合的な問題か、あるいは当該研究のコンセプトそのものに関連した事象か現時点では不明である。

In the current year (Ling and 2004), the contents of the study were published in the first half of the year. In this study, the antibody-Systemized CpG-ODN complex was established. The intracellular protein, antibody, CpG-ODN complex conjugate, intracellular permeability, antibody antibody, intracellular TLR9 antibody, intracellular TLR9 antibody, intracellular protein antibody, specific intracellular protein, specific antibody, antibody It is easier than the description of intracellular proteins (osteoblasts) to make AOC, and to confirm the local distribution of intracellular proteins. Conjugate: the representative cellular bone protein, the β-actin (alpha-tubulin), the AOC, the antibody, the commercial antibody, the pigmented label S, the antibody, the conjugate, the antibody, the antibody. The cell lines (epithelial cell line, hematopoietic cell line, 293T cell line, etc.) were used in this study. After 10 minutes of washing, the intracellular distribution of light pigments was detected by the method of light pigmentation. The results showed that the intracellular signal intensity of light pigments was significantly lower than that of light pigments, and that the intracellular signal intensity of light pigments was significantly lower than that of light pigments. the results showed that the intracellular signal intensity of light pigments was significantly lower than that of light pigments. the results showed that the intracellular signal intensity of light pigments was significantly lower than that of light pigments. the results showed that the intracellular signal intensity of light pigments was significantly lower than that of light pigments. the results showed that the intracellular signal intensity of light pigments was lower than that of light pigments. the results showed that the intracellular signal intensity of light pigments was significantly lower than that of light pigments. The results show that there are many complex problems in the process and technology, and that it is time to study the problems that are not known at any time.