Shigeo Hisamori1,2, Junko Mukohyama3,4,5,6,7,8,9, Sanjay Koul3,4,5,6,7,10, Takanori Hayashi11, Piero Dalerba12,13,14,15,16, Yohei Shimono17,18, Michael Evan Rothenberg1, Masao Maeda11,19, Taichi Isobe1, Luis Enrique Valencia Salazar3,4,5,6,7, Xin Qian1, Darius Michael Johnston1,20, Dalong Qian1, Kaiqin Lao21, Naoya Asai19, Yoshihiro Kakeji8, Vincenzo Alessandro Gennarino22,23,24,25, Debashis Sahoo26. 1. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, 94305, USA. 2. Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, 6068507, Japan. 3. Department of Pathology and Cell Biology, Columbia University, New York, NY, 10032, USA. 4. Department of Medicine (Division of Digestive and Liver Diseases), Columbia University, New York, NY, 10032, USA. 5. Herbert Irving Comprehensive Cancer Center (HICCC), Columbia University, New York, NY, 10032, USA. 6. Digestive and Liver Disease Research Center (DLDRC), Columbia University, New York, NY, 10032, USA. 7. Columbia Stem Cell Initiative (CSCI), Columbia University, New York, NY, 10032, USA. 8. Division of Gastrointestinal Surgery, Kobe University Graduate School of Medicine, Kobe, Hyogo, 6500017, Japan. 9. Department of Hepato-Biliary-Pancreatic and Gastrointestinal Surgery, International University of Health and Welfare (IUHW), Tokyo, 1088329, Japan. 10. Department of Biological Sciences and Geology, Queensboro Community College (QCC), City University of New York (CUNY), New York, NY, 11364, USA. 11. Department of Biochemistry, Fujita Health University School of Medicine, Toyoake, Aichi, 4701192, Japan. 12. Department of Pathology and Cell Biology, Columbia University, New York, NY, 10032, USA. pdd2109@cumc.columbia.edu. 13. Department of Medicine (Division of Digestive and Liver Diseases), Columbia University, New York, NY, 10032, USA. pdd2109@cumc.columbia.edu. 14. Herbert Irving Comprehensive Cancer Center (HICCC), Columbia University, New York, NY, 10032, USA. pdd2109@cumc.columbia.edu. 15. Digestive and Liver Disease Research Center (DLDRC), Columbia University, New York, NY, 10032, USA. pdd2109@cumc.columbia.edu. 16. Columbia Stem Cell Initiative (CSCI), Columbia University, New York, NY, 10032, USA. pdd2109@cumc.columbia.edu. 17. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, 94305, USA. yshimono@fujita-hu.ac.jp. 18. Department of Biochemistry, Fujita Health University School of Medicine, Toyoake, Aichi, 4701192, Japan. yshimono@fujita-hu.ac.jp. 19. Department of Pathology, Fujita Health University School of Medicine, Toyoake, Aichi, 4701192, Japan. 20. Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA, 94305, USA. 21. Genetic Sciences Division (GSD), Thermo Fisher Scientific, South San Francisco, CA, 94080, USA. 22. Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY, 10032, USA. 23. Department of Neurology, Columbia University, New York, NY, 10032, USA. 24. Department of Pediatrics, Columbia University, New York, NY, 10032, USA. 25. Initiative for Columbia Ataxia and Tremor (ICAT), Columbia University Irving Medical Center, New York, NY, 10032, USA. 26. Department of Computer Science and Engineering and Department of Pediatrics, University of California San Diego (UCSD), San Diego, CA, 92123, USA.
Abstract
BACKGROUND: MicroRNAs (miRNAs) are key regulators of stem cell functions, including self-renewal and differentiation. In this study, we aimed to identify miRNAs that are upregulated during terminal differentiation in the human colon epithelium, and elucidate their role in the mechanistic control of stem cell properties. METHODS: "Bottom-of-the-crypt" (EPCAM+/CD44+/CD66alow) and "top-of-the-crypt" (EPCAM+/CD44neg/CD66ahigh) epithelial cells from 8 primary colon specimens (6 human, 2 murine) were purified by flow cytometry and analyzed for differential expression of 335 miRNAs. The miRNAs displaying the highest upregulation in "top-of-the-crypt" (terminally differentiated) epithelial cells were tested for positive correlation and association with survival outcomes in a colon cancer RNA-seq database (n = 439 patients). The two miRNAs with the strongest "top-of-the-crypt" expression profile were evaluated for capacity to downregulate self-renewal effectors and inhibit in vitro proliferation of colon cancer cells, in vitro organoid formation by normal colon epithelial cells and in vivo tumorigenicity by patient-derived xenografts (PDX). RESULTS: Six miRNAs (miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-345) were upregulated in "top-of-the-crypt" cells and positively correlated in expression among colon carcinomas. Overexpression of the three miRNAs with the highest inter-correlation coefficients (miR-200a, miR-200b, miR-200c) associated with improved survival. The top two over-expressed miRNAs (miR-200c, miR-203) cooperated synergistically in suppressing expression of BMI1, a key regulator of self-renewal in stem cell populations, and in inhibiting proliferation, organoid-formation and tumorigenicity of colon epithelial cells. CONCLUSION: In the colon epithelium, terminal differentiation associates with the coordinated upregulation of miR-200c and miR-203, which cooperate to suppress BMI1 and disable the expansion capacity of epithelial cells.
BACKGROUND: MicroRNAs (miRNAs) are key regulators of stem cell functions, including self-renewal and differentiation. In this study, we aimed to identify miRNAs that are upregulated during terminal differentiation in the human colon epithelium, and elucidate their role in the mechanistic control of stem cell properties. METHODS: "Bottom-of-the-crypt" (EPCAM+/CD44+/CD66alow) and "top-of-the-crypt" (EPCAM+/CD44neg/CD66ahigh) epithelial cells from 8 primary colon specimens (6 human, 2 murine) were purified by flow cytometry and analyzed for differential expression of 335 miRNAs. The miRNAs displaying the highest upregulation in "top-of-the-crypt" (terminally differentiated) epithelial cells were tested for positive correlation and association with survival outcomes in a colon cancer RNA-seq database (n = 439 patients). The two miRNAs with the strongest "top-of-the-crypt" expression profile were evaluated for capacity to downregulate self-renewal effectors and inhibit in vitro proliferation of colon cancer cells, in vitro organoid formation by normal colon epithelial cells and in vivo tumorigenicity by patient-derived xenografts (PDX). RESULTS: Six miRNAs (miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-345) were upregulated in "top-of-the-crypt" cells and positively correlated in expression among colon carcinomas. Overexpression of the three miRNAs with the highest inter-correlation coefficients (miR-200a, miR-200b, miR-200c) associated with improved survival. The top two over-expressed miRNAs (miR-200c, miR-203) cooperated synergistically in suppressing expression of BMI1, a key regulator of self-renewal in stem cell populations, and in inhibiting proliferation, organoid-formation and tumorigenicity of colon epithelial cells. CONCLUSION: In the colon epithelium, terminal differentiation associates with the coordinated upregulation of miR-200c and miR-203, which cooperate to suppress BMI1 and disable the expansion capacity of epithelial cells.
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Authors: Nick Barker; Johan H van Es; Jeroen Kuipers; Pekka Kujala; Maaike van den Born; Miranda Cozijnsen; Andrea Haegebarth; Jeroen Korving; Harry Begthel; Peter J Peters; Hans Clevers Journal: Nature Date: 2007-10-14 Impact factor: 49.962
Authors: Piero Dalerba; Tomer Kalisky; Debashis Sahoo; Pradeep S Rajendran; Michael E Rothenberg; Anne A Leyrat; Sopheak Sim; Jennifer Okamoto; Darius M Johnston; Dalong Qian; Maider Zabala; Janet Bueno; Norma F Neff; Jianbin Wang; Andrew A Shelton; Brendan Visser; Shigeo Hisamori; Yohei Shimono; Marc van de Wetering; Hans Clevers; Michael F Clarke; Stephen R Quake Journal: Nat Biotechnol Date: 2011-11-13 Impact factor: 54.908