Transcriptomic Analysis of Breast Cancer Stem Cells and Development of a pALDH1A1:mNeptune Reporter System for Live Tracking

Autor: Violaine Forissier, Marie Winter, Karine Hannebicque, Xuefen Le Bourhis, Samuel Meignan, Amélie Bonnefond, Christophe Ginestier, Mehdi Derhourhi, Ihsan Y El-Sayed, Emmanuelle Charafe-Jauffret, Marie Denoulet, Manon Macario, François Anquez, Sylvain Julien, Nadège Bidan, Justine Bailleul-Dubois, Chann Lagadec, Yukiko T. Matsunaga, Jérémy Duval
Přispěvatelé: Plasticité Cellulaire et Cancer - U908 (CPAC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Institut National de la Santé et de la Recherche Médicale (INSERM), Institut pour la recherche sur le cancer de Lille [Lille] (IRCL), Centre Régional de Lutte contre le Cancer Oscar Lambret [Lille] (UNICANCER/Lille), Université Lille Nord de France (COMUE)-UNICANCER, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Center for International Research on Integrative Biomedical Systems [University of Tokyo] (CIBiS), Institute of Industrial Science (IIS), The University of Tokyo (UTokyo)-The University of Tokyo (UTokyo), Laboratory for Integrated Micro Mechatronics Systems (LIMMS), Centre National de la Recherche Scientifique (CNRS)-The University of Tokyo (UTokyo), Université de Lille, Laboratoire de Physique des Lasers, Atomes et Molécules - UMR 8523 (PhLAM), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Metabolic functional (epi)genomics and molecular mechanisms involved in type 2 diabetes and related diseases - UMR 8199 - UMR 1283 (GI3M), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Lille Nord de France (COMUE), J.B.D. was supported by a grant fellowship from the ARC and IRCL charities. The authors thank the UMR 8199 LIGAN‐PM Genomics platform (Lille, France), which belongs to the 'Federation de Recherche' 3508 Labex EGID (European Genomics Institute for Diabetes, ANR‐10‐LABX‐46) and was supported by the ANR Equipex 2010 session (ANR‐10‐EQPX‐07‐01, 'LIGAN‐PM'). The LIGAN‐PM Genomics platform (Lille, France) is also supported by the FEDER and the Region Hauts de France. This work was supported by the INCa (French Cancer National Institute), Grant number ARC_INCa_LNCC_8068, by Ligue contre le Cancer—Septentrion comittee, by the SIRIC ONCOLille, by 'Cancer du sein—Parlons‐en!' charity 'Pink Ruban' award, and by GEFLUC charity, The authors thank the Flow Cytometry Core facility of the Bio Imaging Center of Lille (BICeL), which is a shared resource of INSERM/CHRU of Lille, and particularly Nathalie Jouy and Emilie Floquet for their help with the FACS experiments., Plasticité Cellulaire et Cancer (CPAC) - U908 (CPAC), Center Oscar Lambret, CRLCC Oscar Lambret, Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU), The University of Tokyo-The University of Tokyo, The University of Tokyo-Centre National de la Recherche Scientifique (CNRS), Génomique Intégrative et Modélisation des Maladies Métaboliques (EGID), Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Université de Lille-UNICANCER, The University of Tokyo (UTokyo)-Centre National de la Recherche Scientifique (CNRS), Metabolic functional (epi)genomics and molecular mechanisms involved in type 2 diabetes and related diseases - UMR 8199 - UMR 1283 (EGENODIA (GI3M))
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Gene isoform
cancer stem cells
[SDV]Life Sciences [q-bio]
Antineoplastic Agents
Breast Neoplasms
[SDV.CAN]Life Sciences [q-bio]/Cancer
Mice
SCID
Biochemistry
Aldehyde Dehydrogenase 1 Family
Transcriptome
fluorescent reporter systems
03 medical and health sciences
Breast cancer
breast cancer
Genes
Reporter
Mice
Inbred NOD
Cancer stem cell
Cell Line
Tumor
Gene expression
Biomarkers
Tumor
Human Umbilical Vein Endothelial Cells
medicine
Animals
Humans
Promoter Regions
Genetic
Molecular Biology
ALDH1A1
030304 developmental biology
0303 health sciences
biology
Genome
Human
Gene Expression Profiling
030302 biochemistry & molecular biology
Reproducibility of Results
Retinal Dehydrogenase
Cellular Reprogramming
medicine.disease
Gene Expression Regulation
Neoplastic
Cell Tracking
Neoplastic Stem Cells
biology.protein
Cancer research
Female
Stem cell
Reprogramming
[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Zdroj: Proteomics
Proteomics, Wiley-VCH Verlag, 2019, 19 (21-22), pp.1800454. ⟨10.1002/pmic.201800454⟩
Proteomics, 2019, 19 (21-22), pp.1800454. ⟨10.1002/pmic.201800454⟩
ISSN: 1615-9853
1615-9861
DOI: 10.1002/pmic.201800454⟩
Popis: International audience; Many solid cancers are hierarchically organized with a small number of cancer stem cells (CSCs) able to regrow a tumor, while their progeny lacks this feature. Breast CSC is known to contribute to therapy resistance. The study of those cells is usually based on their cell-surface markers like CD44high /CD24low/neg or their aldehyde dehydrogenase (ALDH) activity. However, these markers cannot be used to track the dynamics of CSC. Here, a transcriptomic analysis is performed to identify segregating gene expression in CSCs and non-CSCs, sorted by Aldefluor assay. It is observed that among ALDH-associated genes, only ALDH1A1 isoform is increased in CSCs. A CSC reporter system is then developed by using a far red-fluorescent protein (mNeptune) under the control of ALDH1A1 promoter. mNeptune-positive cells exhibit higher sphere-forming capacity, tumor formation, and increased resistance to anticancer therapies. These results indicate that the reporter identifies cells with stemness characteristics. Moreover, live tracking of cells in a microfluidic system reveals a higher extravasation potential of CSCs. Live tracking of non-CSCs under irradiation treatment show, for the first time, live reprogramming of non-CSCs into CSCs. Therefore, the reporter will allow for cell tracking to better understand the implication of CSCs in breast cancer development and recurrence.
Databáze: OpenAIRE
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