title: Monitoring the dynamics of clonal tumour evolution in vivo using secreted luciferases.
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.
authors.affiliation: 1] Molecular Oncology, Philipps-University, 35043 Marburg, Germany [2].
authors.forename: Joël P
authors.initials: JP
authors.lastname: Charles
authors.affiliation: 1] Molecular Oncology, Philipps-University, 35043 Marburg, Germany [2].
authors.forename: Jeannette
authors.initials: J
authors.lastname: Fuchs
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Hefter
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Jonas B
authors.initials: JB
authors.lastname: Vischedyk
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Kleint
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Fotini
authors.initials: F
authors.lastname: Vogiatzi
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Jonas A
authors.initials: JA
authors.lastname: Schäfer
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Nist
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Oleg
authors.initials: O
authors.lastname: Timofeev
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Wanzel
authors.affiliation: Molecular Oncology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Stiewe
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D060965:Clonal Evolution / Q000235:genetics*; D005822:Genetic Vectors; D045325:HCT116 Cells; D006801:Humans; D066298:In Vitro Techniques; D008156:Luciferases; D051379:Mice; D008856:Microscopy, Fluorescence; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D008156:Luciferases