Real-time visualization of intratumoral necrosis using split-luciferase reconstitution by protein trans-splicing.
Real-time visualization of intratumoral necrosis using split-luciferase reconstitution by protein trans-splicing.
复制标题
利用蛋白质转接重组分裂荧光素酶对瘤内坏死进行实时可视化。
DOI：
10.1016/j.omto.2020.12.001
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发表时间：
2021-03-26
期刊：
影响因子：
--
通讯作者：
Hyodo F
中科院分区：
文献类型：
--
作者：
Kagiya G;Sato A;Ogawa R;Hatashita M;Kato M;Kubo M;Kojima F;Kawakami F;Nishimura Y;Abe N;Hyodo F
坏死是细胞死亡的一种形式，不仅随着各种疾病的发展而发生，而且随着肿瘤组织对癌症治疗的反应而发生。因此，通过诱导坏死来寻求癌症治疗的进展可能是根除癌症的最有效方法之一。我们在此描述了一种实时成像系统的发展，以可视化肿瘤内坏死。该系统由表达两种坏死成像报告基因之一的两种类型的细胞组成，所述坏死成像报告基因由连接到两个分裂荧光素酶片段之一的DnaE内含肽序列组成。当在由两种细胞组成的肿瘤中发生坏死时，由于蛋白质反式剪接，两种类型的泄漏报告分子可以重建酶活性，从而在底物存在下发射生物发光。该系统，这是构建与虾衍生的荧光素酶，允许在体外成像的坏死。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，验证了其在体内坏死成像中的应用。因此，所构建的成像系统可能是一个强大的工具，用于优化癌症治疗的治疗条件和评估新的抗癌药物靶向坏死。坏死随着各种疾病的发展而发生，并且发生在对癌症治疗有反应的肿瘤组织中。为了评估不同抗癌药物和放射治疗的临床前有效性，Kagiya及其同事构建了一个实时可视化系统，用于通过蛋白质反式剪接进行分裂荧光素酶重建的肿瘤内坏死。
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 trans-splicing and thereby emit bioluminescence in the presence of the substrate. This system, which was constructed with shrimp-derived luciferase, allowed in vitro imaging of necrosis. We further confirmed real-time imaging of intratumoral necrosis caused by physical or chemical tissue disruption, validating its application in in vivo 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. Necrosis occurs with the development of various diseases and in tumor tissue responding to cancer treatment. To evaluate the preclinical effectiveness of different anticancer drugs and radiation therapy, Kagiya and colleagues have constructed a real-time visualization system for intratumoral necrosis using split-luciferase reconstitution by protein trans-splicing.
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影响因子：
6.6
作者：
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