Genetic Targeting of Organ-Specific Blood Vessels.
Genetic Targeting of Organ-Specific Blood Vessels.
复制标题
器官特异性血管的基因靶向
DOI：
10.1161/circresaha.118.312981
复制
发表时间：
2018-06-22
影响因子：
20.1
通讯作者：
Zhou B
中科院分区：
文献类型：
--
作者：
Pu W;He L;Han X;Tian X;Li Y;Zhang H;Liu Q;Huang X;Zhang L;Wang QD;Yu Z;Yang X;Smart N;Zhou B
原理：人体器官需要血管网络来提供氧气和营养并维持生理功能。不同器官的血管在结构和功能上是异质的。为了更精确地剖析它们在单个器官中的独特体内功能，而不受非位点靶点的潜在干扰，有必要以器官特异性方式遗传靶向它们。目的：本研究的目的是产生一个以器官或组织特异性方式靶向血管内皮细胞的遗传系统，并验证交叉遗传学在体内精确、靶向特异性基因操作中的潜在应用。方法和结果：我们利用两个正交重组系统，Dre-rox和Cre-loxP，建立了一个基于交叉遗传学的遗传靶向系统。使用这种方法，Cre活性仅在同时表达Dre和Cre的细胞中可检测到。应用这个新系统，我们产生了冠状动脉内皮细胞特异性Cre（CoEC-Cre）和脑内皮细胞特异性Cre（BEC-Cre）。通过谱系追踪、基因敲除和过表达实验，我们证明了CoEC-Cre和BEC-Cre分别有效且特异性地靶向心脏和大脑中的血管。通过使用BEC-Cre删除血管内皮生长因子受体2，我们表明血管内皮生长因子信号传导调节中枢神经系统中的血管生成，并且还控制血脑屏障的完整性。结论：我们提供了两个例子来说明交叉遗传学在体内更精确的基因靶向中的应用，即操纵心脏和大脑血管中的基因。更广泛地说，该系统提供了一个有价值的策略，组织特异性基因操作，可广泛应用于其他领域的生物医学研究。
Rationale: Organs of the body require vascular networks to supply oxygen and nutrients and maintain physiological function. The blood vessels of different organs are structurally and functionally heterogeneous in nature. To more precisely dissect their distinct in vivo function in individual organs, without potential interference from off-site targets, it is necessary to genetically target them in an organ-specific manner. Objective: The objective of this study was to generate a genetic system that targets vascular endothelial cells in an organ- or tissue-specific manner and to exemplify the potential application of intersectional genetics for precise, target-specific gene manipulation in vivo. Methods and Results: We took advantage of 2 orthogonal recombination systems, Dre-rox and Cre-loxP, to create a genetic targeting system based on intersectional genetics. Using this approach, Cre activity was only detectable in cells that had expressed both Dre and Cre. Applying this new system, we generated a coronary endothelial cell–specific Cre (CoEC-Cre) and a brain endothelial cell–specific Cre (BEC-Cre). Through lineage tracing, gene knockout and overexpression experiments, we demonstrated that CoEC-Cre and BEC-Cre efficiently and specifically target blood vessels in the heart and brain, respectively. By deletion of vascular endothelial growth factor receptor 2 using BEC-Cre, we showed that vascular endothelial growth factor signaling regulates angiogenesis in the central nervous system and also controls the integrity of the blood-brain barrier. Conclusions: We provide 2 examples to illustrate the use of intersectional genetics for more precise gene targeting in vivo, namely manipulation of genes in blood vessels of the heart and brain. More broadly, this system provides a valuable strategy for tissue-specific gene manipulation that can be widely applied to other fields of biomedical research.