An enhanced bioluminescence-based Annexin V probe for apoptosis detection in vitro and in vivo.
An enhanced bioluminescence-based Annexin V probe for apoptosis detection in vitro and in vivo.
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DOI：
10.1038/cddis.2017.141
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发表时间：
2017-05-25
影响因子：
9
通讯作者：
Daunert S
中科院分区：
文献类型：
--
作者：
Head T;Dau P;Duffort S;Daftarian P;Joshi PM;Vazquez-Padron R;Deo SK;Daunert S
被称为细胞凋亡的受控细胞死亡过程不仅在组织的正常稳态维持中，而且在许多疾病如癌症、神经变性、自身免疫和心血管疾病中具有重要的中心作用。因此，具有选择性检测凋亡细胞的能力的新技术代表了研究这些条件的研究的中心焦点。我们已经开发了一种新的生物传感器，用于检测凋亡细胞，结合有针对性的选择性凋亡细胞膜联蛋白V与生物发光信号产生的灵敏度从血清稳定的突变体海肾荧光素酶（RLuc 8）。我们的数据提供了一个完整的表征这种新的膜联蛋白-海肾融合蛋白（ArFP）的结构和生化特性的构建，以及其在体外检测细胞凋亡的能力的验证。此外，这项工作代表了第一次报告的生物发光膜联蛋白V凋亡传感器在体内使用。有了这个新的结构，我们检查手术诱导的缺血/再灌注，角膜损伤和视网膜细胞死亡作为年龄相关性黄斑变性模型的疾病相关的动物模型内的细胞凋亡。在这些实验中的每一个中，我们证明了ArFP构建体在每种疾病或治疗模型中用于细胞凋亡的检测和生物发光成像的成功应用。ArFP代表了一个重要的新工具，在不断增长的技术试剂盒的凋亡检测，我们的结果在体外和体内实验表明，各种潜在的临床相关的应用，包括癌症治疗筛选和疗效分析，动脉粥样硬化和心血管疾病的检测，以及监测任何数量的其他条件，其中凋亡具有核心作用。
The process of controlled cellular death known as apoptosis has an important central role not only in normal homeostatic maintenance of tissues, but also in numerous diseases such as cancer, neurodegenerative, autoimmune, and cardiovascular diseases. As a result, new technologies with the capability to selectively detect apoptotic cells represent a central focus of research for the study of these conditions. We have developed a new biosensor for the detection of apoptotic cells, incorporating the targeted selectivity for apoptotic cells from Annexin V with the sensitivity of bioluminescence signal generation from a serum-stable mutant of Renilla luciferase (RLuc8). Our data presents a complete characterization of the structural and biochemical properties of this new Annexin-Renilla fusion protein (ArFP) construct, as well as a validation of its ability to detect apoptosis in vitro. Moreover, this work represents the first report of a bioluminescent Annexin V apoptosis sensor utilized in vivo. With this new construct, we examine apoptosis within disease-relevant animal models of surgery-induced ischemia/reperfusion, corneal injury, and retinal cell death as a model of age-related macular degeneration. In each of these experiments, we demonstrate successful application of the ArFP construct for detection and bioluminescence imaging of apoptosis within each disease or treatment model. ArFP represents an important new tool in the continuously growing kit of technologies for apoptosis detection, and our results from both in vitro and in vivo experiments suggest a diverse range of potential clinically relevant applications including cancer therapeutic screening and efficacy analysis, atherosclerosis and cardiovascular disease detection, and the monitoring of any number of other conditions in which apoptosis has a central role.
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DOI：
10.18632/aging.100459
发表时间：
2012-05
期刊：
Aging
影响因子：
--
作者：
Favaloro B;Allocati N;Graziano V;Di Ilio C;De Laurenzi V
通讯作者：
De Laurenzi V
影响因子：
4.8
作者：
Brumatti, Gabriela;Sheridan, Clare;Martin, Seamus J.
通讯作者：
Martin, Seamus J.
DOI：
10.1038/nrm3999
发表时间：
2015-06
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Fuchs Y;Steller H
通讯作者：
Steller H
影响因子：
14.8
作者：
Greenfield, Norma J.
通讯作者：
Greenfield, Norma J.
影响因子：
2.1
作者：
Gabeler, Edward E E;van Hillegersberg, Richard;van Urk, Hero
通讯作者：
van Urk, Hero