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

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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.

Related collections

Most cited references 70

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A long-sought milestone in microfluidics research has been the development of integrated technology for scalable analysis of transcription in single cells. Here we present a fully integrated microfluidic device capable of performing high-precision RT-qPCR measurements of gene expression from hundreds of single cells per run. Our device executes all steps of single-cell processing, including cell capture, cell lysis, reverse transcription, and quantitative PCR. In addition to higher throughput and reduced cost, we show that nanoliter volume processing reduced measurement noise, increased sensitivity, and provided single nucleotide specificity. We apply this technology to 3,300 single-cell measurements of (i) miRNA expression in K562 cells, (ii) coregulation of a miRNA and one of its target transcripts during differentiation in embryonic stem cells, and (iii) single nucleotide variant detection in primary lobular breast cancer cells. The core functionality established here provides the foundation from which a variety of on-chip single-cell transcription analyses will be developed.

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Author and article information

Journal

Journal ID (nlm-journal-id): 101637426

Journal ID (pubmed-jr-id): 42917

Journal ID (nlm-ta): Biotechnol Rep (Amst)

Journal ID (iso-abbrev): Biotechnol Rep (Amst)

Title: Biotechnology reports (Amsterdam, Netherlands)

ISSN (Electronic): 2215-017X

Publication date Nihms-submitted: 12 November 2015

Publication date (Electronic): 1 August 2015

Publication date (Print): September 2015

Publication date PMC-release: 15 March 2016

Volume: 7

Pages: 157-165

Affiliations

[a ] Division of Neurointerventional Radiology, Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA, USA

[b ] Department of Anatomy, University of California, San Francisco, CA, USA

[c ] Division of Experimental Medicine, Department of Medicine, University of California, San Francisco, CA, USA

[d ] Center for Cerebrovascular Research, Department of Anesthesia and Perioperative Care, University of California, San Francisco, CA, USA

[e ] Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan

Author notes

[* ] Corresponding author at: Interventional Neuroradiology, 505 Parnassus Avenue, Box-0628, San Francisco, CA 94143, USA. Fax: +1 415 353 8606. cooke@ 123456ucsf.edu (D.L. Cooke).

[1]

Co-senior authors.

Article

Manuscript ID: NIHMS736802

DOI: 10.1016/j.btre.2015.07.001

PMC ID: 4792280

PubMed ID: 26989654

SO-VID: fcb7d089-f165-483e-80c0-0813a1321bd4

License:

This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/).

^✩ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.