Intracellular autofluorescence: a biomarker for epithelial cancer stem cells

Irene Miranda-Lorenzo; Jorge Dorado; Enza Lonardo; Sonia Alcala; Alicia G Serrano; Jenifer Clausell-Tormos; Michele Cioffi; Diego Megias; Sladjana Zagorac; Anamaria Balic; Manuel Hidalgo; Mert Erkan; Joerg Kleeff; Aldo Scarpa; Bruno Sainz Jr; Christopher Heeschen

doi:10.1038/nmeth.3112

Intracellular autofluorescence: a biomarker for epithelial cancer stem cells

Nat Methods. 2014 Nov;11(11):1161-9. doi: 10.1038/nmeth.3112. Epub 2014 Sep 28.

Authors

Irene Miranda-Lorenzo¹, Jorge Dorado¹, Enza Lonardo¹, Sonia Alcala¹, Alicia G Serrano², Jenifer Clausell-Tormos¹, Michele Cioffi¹, Diego Megias³, Sladjana Zagorac¹, Anamaria Balic¹, Manuel Hidalgo⁴, Mert Erkan⁵, Joerg Kleeff⁵, Aldo Scarpa⁶, Bruno Sainz Jr¹, Christopher Heeschen⁷

Affiliations

¹ Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain.
² Melanoma Group, Molecular Pathology Programme, CNIO, Madrid, Spain.
³ Confocal Microscopy Unit, Biotechnology Programme, CNIO, Madrid, Spain.
⁴ Gastrointestinal Cancer Clinical Research Unit, Clinical Research Programme, CNIO, Madrid, Spain.
⁵ Chirurgische Klinik, Technische Universität München, Munich, Germany.
⁶ Università degli Studi di Verona, Centro Ricerca Applicata ARC-NET, Verona, Italy.
⁷ 1] Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. [2] Centre for Stem Cells in Cancer &Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK.

PMID: 25262208
DOI: 10.1038/nmeth.3112

Abstract

Cancer stem cells (CSCs) are thought to drive tumor growth, metastasis and chemoresistance. Although surface markers such as CD133 and CD44 have been successfully used to isolate CSCs, their expression is not exclusively linked to the CSC phenotype and is prone to environmental alteration. We identified cells with an autofluorescent subcellular compartment that exclusively showed CSC features across different human tumor types. Primary tumor-derived autofluorescent cells did not overlap with side-population (SP) cells, were enriched in sphere culture and during chemotherapy, strongly expressed pluripotency-associated genes, were highly metastatic and showed long-term in vivo tumorigenicity, even at the single-cell level. Autofluorescence was due to riboflavin accumulation in membrane-bounded cytoplasmic structures bearing ATP-dependent ABCG2 transporters. In summary, we identified and characterized an intrinsic autofluorescent phenotype in CSCs of diverse epithelial cancers and used this marker to isolate and characterize these cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters / metabolism
Animals
Autophagy
Autophagy-Related Protein 12
Biomarkers, Tumor / metabolism*
Carcinoma, Hepatocellular / pathology
Carcinoma, Non-Small-Cell Lung / pathology
Carcinoma, Pancreatic Ductal / pathology
Cell Separation / methods*
Colorectal Neoplasms / pathology
Female
Humans
Liver Neoplasms / pathology
Lung Neoplasms / pathology
Mice
Mice, Nude
Microtubule-Associated Proteins / biosynthesis
Neoplasm Proteins / metabolism
Neoplastic Stem Cells / metabolism*
Neoplastic Stem Cells / pathology*
Optical Imaging / methods*
Pancreatic Neoplasms / pathology
Riboflavin / metabolism*
Small Ubiquitin-Related Modifier Proteins / biosynthesis
Tumor Cells, Cultured

Substances

ABCG2 protein, human
ATG12 protein, human
ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters
Autophagy-Related Protein 12
Biomarkers, Tumor
MAP1LC3A protein, human
Microtubule-Associated Proteins
Neoplasm Proteins
Small Ubiquitin-Related Modifier Proteins
Riboflavin