Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels✩

Zhengda Sun; Devon A Lawson; Elizabeth Sinclair; Chih-Yang Wang; Ming-Derg Lai; Steven W Hetts; Randall T Higashida; Christopher F Dowd; Van V Halbach; Zena Werb; Hua Su; Daniel L Cooke

doi:10.1016/j.btre.2015.07.001

Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels^✩

Biotechnol Rep (Amst). 2015 Sep:7:157-165. doi: 10.1016/j.btre.2015.07.001. Epub 2015 Aug 1.

Authors

Affiliations

¹ Division of Neurointerventional Radiology, Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA, USA.
² Department of Anatomy, University of California, San Francisco, CA, USA.
³ Division of Experimental Medicine, Department of Medicine, University of California, San Francisco, CA, USA.
⁴ Department of Anatomy, University of California, San Francisco, CA, USA; Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁵ Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁶ Center for Cerebrovascular Research, Department of Anesthesia and Perioperative Care, University of California, San Francisco, CA, USA.

Abstract

Purpose: To develop a strategy of achieving targeted collection of endothelial cells (ECs) by endovascular methods and analyzing the gene expression profiles of collected single ECs.

Methods and results: 134 ECs and 37 leukocytes were collected from four patients' intra-iliac artery endovascular guide wires by fluorescence activated cell sorting (FACS) and analyzed by single-cell quantitative RT-PCR for expression profile of 48 genes. Compared to CD45⁺ leukocytes, the ECs expressed higher levels (p < 0.05) of EC surface markers used on FACS and other EC related genes. The gene expression profile showed that these isolated ECs fell into two clusters, A and B, that differentially expressed 19 genes related to angiogenesis, inflammation and extracellular matrix remodeling, with cluster B ECs have demonstrating similarities to senescent or aging ECs.

Conclusion: Combination of endovascular device sampling, FACS and single-cell quantitative RT-PCR is a feasible method for analyzing EC gene expression profile in vascular lesions.

Keywords: Gene expression of artery endothelial cells; Single cell quantitative RT-PCR; Targeted endothelial cell sampling.

Abstract

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