Sequential cancer mutations in cultured human intestinal stem cells
| Autor: | Gerald Schwank, Arjan Buijs, René M. Overmeer, Norman Sachs, Hugo J. Snippert, Cheryl Zimberlin, Ruben van Boxtel, Harry Begthel, Jeroen Korving, Richard H. van Jaarsveld, Marc van de Wetering, Hans Clevers, Jarno Drost, Meike Logtenberg, Jan Paul Medema, Geert J. P. L. Kops, Edwin Cuppen, G. Johan A. Offerhaus, Bas Ponsioen |
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| Přispěvatelé: | Radiotherapy, Other departments, CCA -Cancer Center Amsterdam, Center of Experimental and Molecular Medicine, Hubrecht Institute for Developmental Biology and Stem Cell Research |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
HUMAN COLON
medicine.medical_treatment medicine.disease_cause SMAD4 Piperazines COLORECTAL-CANCER Mice Epidermal growth factor Intestinal Mucosa Stem Cell Niche Child Non-U.S. Gov't Smad4 Protein Genetics Multidisciplinary Stem Cells Research Support Non-U.S. Gov't Imidazoles Wnt signaling pathway Middle Aged Intestines Child Preschool Heterografts Intercellular Signaling Peptides and Proteins Female KRAS Stem cell Colorectal Neoplasms STRUCTURAL BASIS Genes APC Xenotransplantation EPITHELIUM Mutagenesis (molecular biology technique) Biology Research Support Proto-Oncogene Proteins p21(ras) ORGANOIDS ADENOMAS REVEALS medicine Organoid Journal Article Animals Humans Neoplasm Invasiveness Noggin IN-VITRO EXPANSION Aneuploidy Genes p53 TRANSFORMATION Mutation Mutagenesis Site-Directed Cancer research CRISPR-Cas Systems Neoplasm Transplantation |
| Zdroj: | Nature, 521(7550), 43-37. Nature Publishing Group Nature, 521(7550), 43-7. Nature Publishing Group Nature, 521(7550), 43. Nature Publishing Group |
| ISSN: | 0028-0836 |
| Popis: | Crypt stem cells represent the cells of origin for intestinal neoplasia. Both mouse and human intestinal stem cells can be cultured in medium containing the stem-cell-niche factors WNT, R-spondin, epidermal growth factor (EGF) and noggin over long time periods as epithelial organoids that remain genetically and phenotypically stable. Here we utilize CRISPR/Cas9 technology for targeted gene modification of four of the most commonly mutated colorectal cancer genes (APC, P53 (also known as TP53), KRAS and SMAD4) in cultured human intestinal stem cells. Mutant organoids can be selected by removing individual growth factors from the culture medium. Quadruple mutants grow independently of all stem-cell-niche factors and tolerate the presence of the P53 stabilizer nutlin-3. Upon xenotransplantation into mice, quadruple mutants grow as tumours with features of invasive carcinoma. Finally, combined loss of APC and P53 is sufficient for the appearance of extensive aneuploidy, a hallmark of tumour progression. |
| Databáze: | OpenAIRE |
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