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Monitoring the dynamics of clonal tumour evolution in vivo using secreted luciferases
Joël P Charles1, Jeannette Fuchs1, Mirjam Hefter2
11] Molecular Oncology, Philipps-University, 35043 Marburg, Germany [2].
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.
Insights
Researchers developed a novel toolkit using secreted luciferases to track tumor cell clones. This minimally invasive method accurately quantifies the dynamic evolution of distinct tumor subclones in preclinical models.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Tumors exhibit heterogeneity and clonal evolution during progression, metastasis, and therapy response.
- Short hairpin RNAs (shRNAs) are used to study genetic alterations but tracking methods are limited.
- Fluorescent reporters are unsuitable for solid tumors in vivo, hindering clonal evolution studies.
Purpose of the Study:
- To develop a novel toolkit for tracking multiple shRNA-expressing tumor cell clones.
- To enable minimally invasive, in vivo quantification of clonal evolution in solid tumors.
- To provide a versatile tool for preclinical cancer research.
Main Methods:
- Development of a toolkit utilizing secreted luciferases.
- Competitive in vitro and in vivo tracking of two distinct shRNA-expressing tumor cell clones.
- Measurement of secreted luciferase activity in the bloodstream of tumor-bearing mice.
Main Results:
- Robust measurement of secreted luciferase activity in mouse blood.
- Accurate, minimally invasive quantification of dynamic tumor subclone evolution.
- Successful application in preclinical models of tumor development, metastasis, and therapy.
Conclusions:
- Secreted luciferases offer a powerful method for tracking tumor cell clones in vivo.
- This toolkit overcomes limitations of fluorescent reporters for solid tumor studies.
- The approach facilitates detailed analysis of clonal dynamics in cancer progression and treatment.
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