Self-sustaining long-term 3D epithelioid cultures reveal drivers of clonal expansion in esophageal epithelium
Aging epithelia are colonized by somatic mutations, which are subjected to selection influenced by intrinsic and extrinsic factors. The lack of suitable culture systems has slowed the study of this and other long-term biological processes. Here, we describe epithelioids, a facile, cost-effective method of culturing multiple mouse and human epithelia. Esophageal epithelioids self-maintain without passaging for at least 1 year, maintaining a three-dimensional structure with proliferative basal cells that differentiate into suprabasal cells, which eventually shed and retain genomic stability. Live imaging over 5 months showed that epithelioids replicate in vivo cell dynamics. Epithelioids support genetic manipulation and enable the study of mutant cell competition and selection in three-dimensional epithelia, and show how anti-cancer treatments modulate competition between transformed and wild-type cells. Finally, a targeted CRISPR-Cas9 screen shows that epithelioids recapitulate mutant gene selection in aging human esophagus and identifies additional drivers of clonal expansion, resolving the genetic networks underpinning competitive fitness.
Authors
- Moritz Gerstung (ORCID: https://orcid.org/0000-0001-6709-963X)
- Charlotte King (ORCID: https://orcid.org/0000-0003-2543-1993)
- Alan R. Lowe (ORCID: https://orcid.org/0000-0002-0558-3597)
- Maria P. Alcolea (ORCID: https://orcid.org/0000-0002-8396-6454)
- Argyro Kalogeropoulou
- Swee Hoe Ong (ORCID: https://orcid.org/0000-0002-3629-5387)
- Gabriel Piedrafita (ORCID: https://orcid.org/0000-0001-8701-1084)
- Krishnaa T. A. Mahbubani (ORCID: https://orcid.org/0000-0002-1327-2334)
- Bartomeu Colom (ORCID: https://orcid.org/0000-0001-6256-6462)
- Philip H. Jones (ORCID: https://orcid.org/0000-0002-5904-795X)
- Albert Herms (ORCID: https://orcid.org/0000-0003-2999-8196)
- Kourosh Saeb‐Parsy (ORCID: https://orcid.org/0000-0002-0633-3696)
- Maria Teresa Béjar (ORCID: https://orcid.org/0000-0002-8850-0178)
- Jose Antonio Valverde-Lopez
- Ujjwal Banerjee
- Stefan C. Dentro (ORCID: https://orcid.org/0000-0002-0478-9729)
- Emilie Abby (ORCID: https://orcid.org/0000-0003-0284-4760)
- David Fernández‐Antorán (ORCID: https://orcid.org/0000-0003-1608-570X)
- I. Caseda
- Inês Sofia dos Santos Rodrigues Ferreira (ORCID: https://orcid.org/0000-0003-2937-7728)
- Kasumi Murai
- Sara Vidal-Notari (ORCID: https://orcid.org/0009-0002-7741-4976)
Institutions
- Universidad Complutense de Madrid (ES)
- European Bioinformatics Institute (GB)
- Wellcome/MRC Cambridge Stem Cell Institute (GB)
- University of Cambridge (GB)
- The Gurdon Institute (GB)
- Wellcome Sanger Institute (GB)
- National Institute for Health and Care Research (GB)
- Fundacion Agencia Aragonesa para la Investigacion y el Desarrollo (ES)
- Spanish National Cancer Research Centre (ES)
- Hutchison/MRC Research Centre (GB)
- DKFZ-ZMBH Alliance (DE)
- Institute of Structural and Molecular Biology (GB)
- Consorci Institut D'Investigacions Biomediques August Pi I Sunyer (ES)
- University College London (GB)
- Universitat de Barcelona (ES)
Publication Details
- Journal
- Nature Genetics
- Published
- 2024-09-23
- DOI
- https://doi.org/10.1038/s41588-024-01875-8
- Citations
- 12
- Primary Topic
- Tissue Engineering and Regenerative Medicine
- Type
- article
- Field-Weighted Citation Impact
- 6.87
Funders
- European Molecular Biology Organization
- Wellcome
- Worldwide Cancer Research
- Cancer Research UK
- Leverhulme Trust
- Royal Society
- Agencia Canaria de Investigación, Innovación y Sociedad de la Información
- National Centre for the Replacement Refinement and Reduction of Animals in Research
- Medical Research Council
- FP7 Nuclear Fission, Safety and Radiation Protection