Genetic Targeting of Organ-Specific Blood Vessels

Wenjuan Pu; Lingjuan He; Ximeng Han; Xueying Tian; Yan Li; Hui Zhang; Qiaozhen Liu; Xiuzhen Huang; Libo Zhang; Qing-Dong Wang; Zhenyang Yu; Xiao Yang; Nicola Smart; Bin Zhou

doi:10.1161/CIRCRESAHA.118.312981

Genetic Targeting of Organ-Specific Blood Vessels

Circ Res. 2018 Jun 22;123(1):86-99. doi: 10.1161/CIRCRESAHA.118.312981. Epub 2018 May 15.

Authors

Wenjuan Pu^{1

2}, Lingjuan He^{1

2}, Ximeng Han³, Xueying Tian^{1

2}, Yan Li^{1

2}, Hui Zhang^{1

3}, Qiaozhen Liu^{1

2}, Xiuzhen Huang^{1

2}, Libo Zhang^{1

2}, Qing-Dong Wang⁴, Zhenyang Yu⁵, Xiao Yang⁵, Nicola Smart⁶, Bin Zhou^{7

2

3

8}

Affiliations

¹ From the State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences (W.P., L.H., X.T., Y.L., H.Z., Q.L., X. Huang, L.Z., B.Z.).
² Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences (W.P., L.H., X.T., Y.L., Q.L., X. Huang, L.Z., B.Z.).
³ Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; School of Life Science and Technology, Shanghai Tech University, China (X. Han, H.Z., B.Z.).
⁴ Bioscience Heart Failure, Cardiovascular and Metabolic Diseases, IMED Biotech Unit, AstraZeneca, Gothenburg, Sweden (Q.-D.W.).
⁵ State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Laboratory of Proteomics, Institute of Biotechnology, China (Z.Y., X.Y.).
⁶ British Heart Foundation Centre of Regenerative Medicine, Department of Physiology, Anatomy and Genetics, University of Oxford, United Kingdom (N.S.).
⁷ From the State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences (W.P., L.H., X.T., Y.L., H.Z., Q.L., X. Huang, L.Z., B.Z.) zhoubin@sibs.ac.cn.
⁸ Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou, China (B.Z.).

Abstract

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.

Keywords: blood vessels; blood-brain barrier; endothelial cell; models, animal; recombination, genetic.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blood Vessels*
Blood-Brain Barrier
Brain / blood supply*
Cell Hypoxia
Coronary Vessels*
Endothelial Cells
Gene Knockout Techniques
Gene Targeting / methods*
In Situ Hybridization / methods
Mice
Neovascularization, Physiologic
Organ Specificity
Receptors, Vascular Endothelial Growth Factor / physiology
Vascular Endothelial Growth Factor A / physiology

Substances

Vascular Endothelial Growth Factor A
Receptors, Vascular Endothelial Growth Factor

Abstract

Publication types

MeSH terms

Substances

Grants and funding