Monitoring the dynamics of clonal tumour evolution in vivo using secreted luciferases

Joël P. Charles, Jeannette Fuchs, Mirjam Hefter, Jonas B. Vischedyk, Maximilian Kleint, Fotini Vogiatzi, Jonas A. Schäfer, Andrea Nist, Oleg Timofeev, Michael Wanzel and Thorsten Stiewe ()
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Joël P. Charles: Molecular Oncology, Philipps-University
Jeannette Fuchs: Molecular Oncology, Philipps-University
Mirjam Hefter: Molecular Oncology, Philipps-University
Jonas B. Vischedyk: Molecular Oncology, Philipps-University
Maximilian Kleint: Molecular Oncology, Philipps-University
Fotini Vogiatzi: Molecular Oncology, Philipps-University
Jonas A. Schäfer: Molecular Oncology, Philipps-University
Andrea Nist: Molecular Oncology, Philipps-University
Oleg Timofeev: Molecular Oncology, Philipps-University
Michael Wanzel: Molecular Oncology, Philipps-University
Thorsten Stiewe: Molecular Oncology, Philipps-University

Nature Communications, 2014, vol. 5, issue 1, 1-11

Abstract: Abstract Tumours are heterogeneous cell populations that undergo clonal evolution during tumour progression, metastasis and response to therapy. Short hairpin RNAs (shRNAs) generate stable loss-of-function phenotypes and are versatile experimental tools to explore the contribution of individual genetic alterations to clonal evolution. In these experiments tumour cells carrying shRNAs are commonly tracked with fluorescent reporters. While this works well for cell culture studies and leukaemia mouse models, fluorescent reporters are poorly suited for animals with solid tumours—the most common tumour types in cancer patients. Here we develop a toolkit that uses secreted luciferases to track the fate of two different shRNA-expressing tumour cell clones competitively, both in vitro and in vivo. We demonstrate that secreted luciferase activities can be measured robustly in the blood stream of tumour-bearing mice to accurately quantify, in a minimally invasive manner, the dynamic evolution of two genetically distinct tumour subclones in preclinical mouse models of tumour development, metastasis and therapy.

Date: 2014
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DOI: 10.1038/ncomms4981

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