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Title: Longitudinal Monitoring of Intra-Tumoural Heterogeneity Using Optical Barcoding of Patient-Derived Colorectal Tumour Models.
Austin Authors: Shembrey, Carolyn;Smith, Jai;Grandin, Mélodie;Williams, Nathalia;Cho, Hyun-Jung;Mølck, Christina;Behrenbruch, Corina;Thomson, Benjamin Nj;Heriot, Alexander G;Merino, Delphine;Hollande, Frédéric
Affiliation: Department of Surgery, St Vincent's Hospital, Melbourne, VIC 3065, Australia..
Department of Medical Biology, The Faculty of Medicine, Dentistry and Health Science, University of Melbourne, Melbourne, VIC 3010, Australia..
Biological Optical Microscopy Platform, University of Melbourne, Melbourne, VIC 3010, Australia..
Department of General Surgical Specialties, The Royal Melbourne Hospital, University of Melbourne, Melbourne, VIC 3050, Australia..
Department of Surgery, the Royal Melbourne Hospital, University of Melbourne, Melbourne, VIC 3050, Australia..
The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC 3010, Australia..
Department of Cancer Surgery, Peter MacCallum Cancer Centre, Melbourne, VIC 3000, Australia..
Department of Clinical Pathology, University of Melbourne, Melbourne, VIC 3000, Australia..
Victorian Comprehensive Cancer Centre, University of Melbourne Centre for Cancer Research, Melbourne, VIC 3000, Australia..
Olivia Newton-John Cancer Research Institute
School of Cancer Medicine, La Trobe University, Melbourne, VIC 3086, Australia..
Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Melbourne, VIC 3052, Australia..
Issue Date: 24-Jan-2022 2022
Publication information: Cancers 2022; 14(3): 581
Abstract: Geno- and phenotypic heterogeneity amongst cancer cell subpopulations are established drivers of treatment resistance and tumour recurrence. However, due to the technical difficulty associated with studying such intra-tumoural heterogeneity, this phenomenon is seldom interrogated in conventional cell culture models. Here, we employ a fluorescent lineage technique termed "optical barcoding" (OBC) to perform simultaneous longitudinal tracking of spatio-temporal fate in 64 patient-derived colorectal cancer subclones. To do so, patient-derived cancer cell lines and organoids were labelled with discrete combinations of reporter constructs, stably integrated into the genome and thus passed on from the founder cell to all its clonal descendants. This strategy enables the longitudinal monitoring of individual cell lineages based upon their unique optical barcodes. By designing a novel panel of six fluorescent proteins, the maximum theoretical subpopulation resolution of 64 discriminable subpopulations was achieved, greatly improving throughput compared with previous studies. We demonstrate that all subpopulations can be purified from complex clonal mixtures via flow cytometry, permitting the downstream isolation and analysis of any lineages of interest. Moreover, we outline an optimized imaging protocol that can be used to image optical barcodes in real-time, allowing for clonal dynamics to be resolved in live cells. In contrast with the limited intra-tumour heterogeneity observed in conventional 2D cell lines, the OBC technique was successfully used to quantify dynamic clonal expansions and contractions in 3D patient-derived organoids, which were previously demonstrated to better recapitulate the heterogeneity of their parental tumour material. In summary, we present OBC as a user-friendly, inexpensive, and high-throughput technique for monitoring intra-tumoural heterogeneity in in vitro cell culture models.
DOI: 10.3390/cancers14030581
ORCID: 0000-0003-4997-6189
Journal: Cancers
PubMed URL: 35158849
PubMed URL:
ISSN: 2072-6694
Type: Journal Article
Subjects: cell culture techniques
cell lineage
clonal evolution
colorectal neoplasms
longitudinal imaging
neoplasm recurrence
tumour heterogeneity
Appears in Collections:Journal articles

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