Fluorometric Quantification of Single-Cell Velocities to Investigate Cancer Metastasis

Erin Elizabeth Edwards; Katherine Gayle Birmingham; Meghan Jeanne O'Melia; Jaeho Oh; Susan Napier Thomas

doi:10.1016/j.cels.2018.10.005

Fluorometric Quantification of Single-Cell Velocities to Investigate Cancer Metastasis

Cell Syst. 2018 Nov 28;7(5):496-509.e6. doi: 10.1016/j.cels.2018.10.005. Epub 2018 Nov 7.

Authors

Erin Elizabeth Edwards¹, Katherine Gayle Birmingham², Meghan Jeanne O'Melia³, Jaeho Oh⁴, Susan Napier Thomas⁵

Affiliations

¹ Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
² Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA; George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.
³ Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
⁴ George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.
⁵ Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA; George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA; Winship Cancer Institute, Emory University, Atlanta, GA 30322, USA. Electronic address: susan.thomas@gatech.edu.

Abstract

Hematogenous metastasis is a multistep, selectin-regulated process whose mechanisms remain poorly understood. To investigate this biological pathway of cancer dissemination and better understand circulating cancer cells, we developed a high-throughput methodology that integrates organ-on-chip-like microfluidic and photoconvertible protein technologies. Our approach can ascribe single-cell velocity as a traceable cell property for off-chip analysis of the direct relationships between cell molecular profiles and adhesive phenotypes in the context of physiologically relevant fluid flow. We interrogate how natively expressed selectin ligands relate to colon cancer cell rolling frequencies and velocities and provide context for previously reported disparities in in vitro and in vivo models of selectin-mediated adhesion and metastasis. This integrated methodology represents a versatile approach for the development of anti-metastatic therapeutics as well as to generate and test mechanistic hypotheses regarding spatiotemporal processes that occur over timescales of seconds to hours with single-cell resolution.

Keywords: CD44; carcinoembryonic antigen; colon cancer; flow cytometry; high-throughput; microfluidic; photoconvertible; selectin; sialyl Lewis x; single-cell.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Line, Tumor
Colonic Neoplasms / pathology*
Female
Fluorometry / methods*
Humans
Neoplasm Metastasis*
Neoplastic Cells, Circulating*
Xenograft Model Antitumor Assays

Abstract

Publication types

MeSH terms

Grants and funding