Kagiya Go, Sato Ayaka, Ogawa Ryohei, Hatashita Masanori, Kato Mana, Kubo Makoto, Kojima Fumiaki, Kawakami Fumitaka, Nishimura Yukari, Abe Naoya, Hyodo Fuminori
School of Allied Health Sciences, Kitasato University, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa 252-0373, Japan.
Regenerative Medicine and Cell Design Research Facility, Kitasato University, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa 252-0373, Japan.
Mol Ther Oncolytics. 2020 Dec 10;20:48-58. doi: 10.1016/j.omto.2020.12.001. eCollection 2021 Mar 26.
Necrosis, a form of cell death, occurs not only with the development of various diseases but also with a tumor tissue response to cancer treatment. Therefore, pursuing progress for cancer therapy through induction of necrosis may be one of the most effective approaches for cancer eradication. We herein describe the development of a real-time imaging system to visualize intratumoral necrosis. The system is composed of two types of cells expressing either one of two necrosis imaging reporters that consist of a DnaE intein sequence linking to one of two split-luciferase fragments. When necrosis occurs in a tumor composed of both of the cells, the two types of leaked reporters can reconstitute the enzymatic activity as a result of protein -splicing and thereby emit bioluminescence in the presence of the substrate. This system, which was constructed with shrimp-derived luciferase, allowed imaging of necrosis. We further confirmed real-time imaging of intratumoral necrosis caused by physical or chemical tissue disruption, validating its application in necrosis imaging. Thus, the constructed imaging system could be a powerful tool for the optimization of the therapeutic condition for cancer therapy and for the evaluation of novel anticancer drugs targeting necrosis.
坏死是一种细胞死亡形式，不仅在各种疾病的发展过程中出现，也会在肿瘤组织对癌症治疗的反应中发生。因此，通过诱导坏死来推动癌症治疗进展可能是根除癌症最有效的方法之一。我们在此描述了一种用于可视化肿瘤内坏死的实时成像系统的开发。该系统由两种细胞组成，这两种细胞分别表达两种坏死成像报告基因中的一种，这两种报告基因由与两个分裂荧光素酶片段之一相连的DnaE内含肽序列组成。当由这两种细胞组成的肿瘤中发生坏死时，两种类型的泄漏报告基因可由于蛋白质剪接而重新构成酶活性，从而在有底物存在时发出生物发光。这个用虾源荧光素酶构建的系统能够对坏死进行成像。我们进一步证实了由物理或化学组织破坏引起的肿瘤内坏死的实时成像，验证了其在坏死成像中的应用。因此，构建的成像系统可能是优化癌症治疗条件和评估靶向坏死的新型抗癌药物的有力工具。
