Real-time visualization of intratumoral necrosis using split-luciferase reconstitution by protein trans-splicing
| Autor: | Mana Kato, Fumitaka Kawakami, Fumiaki Kojima, Ryohei Ogawa, Fuminori Hyodo, Makoto Kubo, Naoya Abe, Masanori Hatashita, Yukari Nishimura, Ayaka Sato, Go Kagiya |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Cancer Research Programmed cell death Necrosis dnaE Trans-splicing intein lcsh:RC254-282 necrosis extein 03 medical and health sciences 0302 clinical medicine split-luciferase reconstitution medicine Bioluminescence Pharmacology (medical) Luciferase bioluminescent imaging Chemistry Cancer lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens medicine.disease cell death 030104 developmental biology Oncology 030220 oncology & carcinogenesis Cancer research Molecular Medicine Original Article protein trans-splicing medicine.symptom Intein necrosis imaging reporter |
| Zdroj: | Molecular Therapy Oncolytics Molecular Therapy: Oncolytics, Vol 20, Iss, Pp 48-58 (2021) |
| ISSN: | 2372-7705 |
| DOI: | 10.1016/j.omto.2020.12.001 |
| Popis: | 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. Graphical Abstract 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. |
| Databáze: | OpenAIRE |
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