抄録
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-derivedxenografts (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.
| 本文言語 | English |
|---|---|
| ページ（範囲） | 407-422 |
| ページ数 | 16 |
| ジャーナル | Journal of Gastroenterology |
| 巻 | 57 |
| 号 | 6 |
| DOI | |
| 出版ステータス | Published - 06-2022 |
All Science Journal Classification (ASJC) codes
- 消化器病学
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Upregulation of BMI1-suppressor miRNAs (miR-200c, miR-203) during terminal differentiation of colon epithelial cells. / Hisamori, Shigeo; Mukohyama, Junko; Koul, Sanjay その他.
In: Journal of Gastroenterology, Vol. 57, No. 6, 06.2022, p. 407-422.研究成果: Article › 査読
TY - JOUR
T1 - Upregulation of BMI1-suppressor miRNAs (miR-200c, miR-203) during terminal differentiation of colon epithelial cells
AU - Hisamori, Shigeo
AU - Mukohyama, Junko
AU - Koul, Sanjay
AU - Hayashi, Takanori
AU - Rothenberg, Michael Evan
AU - Maeda, Masao
AU - Isobe, Taichi
AU - Valencia Salazar, Luis Enrique
AU - Qian, Xin
AU - Johnston, Darius Michael
AU - Qian, Dalong
AU - Lao, Kaiqin
AU - Asai, Naoya
AU - Kakeji, Yoshihiro
AU - Gennarino, Vincenzo Alessandro
AU - Sahoo, Debashis
AU - Dalerba, Piero
AU - Shimono, Yohei
N1 - Funding Information: The authors wish to thank Michael F. Clarke, MD (Institute for Stem Cell Biology and Regenerative Medicine , Stanford University) for his continuing mentorship and invaluable scientific insights. We thank Dr. Seetha V. Srinivasan (Herbert Irving Comprehensive Cancer Center , Columbia University) for editorial assistance during the preparation of this manuscript. We thank Yusuke Akama and Hiromi Yamazaki (TechnoPro Inc., Fujita Health University) and Hiroaki Sakai (Fujita Health University) for exceptional technical assistance. This work was supported by: (1) the Japan Society for the Promotion of Science (JSPS), through a Research Fellowship for Young Scientists (to Shigeo Hisamori), an Overseas Research Fellowship (to Junko Mukohyama) and the Grants-in-Aid for Scientific Research (KAKENHI) program, with specific regard to grants 17K16555 (to J.M.), 15K14381 (to Yohei Shimono), 18K07231 (to Y.S.), 19K09106 (to Y.S.) and 21H02769 (to Y.S.); (2) the Japan-Belgium Research Cooperative Program (to Y.S.); (3) a post-doctoral fellowship from the Uehara Memorial Foundation (to J.M.); (4) a post-doctoral scholarship from The Cell Science Research Foundation (to J.M.); (5) the Empire State Institutional Training Program (DOH01-C30291GG-3450000) of the New York State Stem Cell Science (NYSTEM) agency (to J.M.); (6) a grant from the Princess Takamatsu Cancer Research Fund (to Y.S.); (7) a Grant from Fujita Health University (to Y.S.); (8) an Extramural Collaborative Research Grant from the Cancer Research Institute of Kanazawa University (to Y.S.); (9) a Grant from the Japan Association for Development of Community Medicine (to T.I.); (10) a BD Biosciences 2011 Stem Cell Research Grant (to Piero Dalerba); (11) a 2016 Runyon-Rachleff Innovator Award (DRR-44-16) from the Damon Runyon Cancer Research Foundation (to P.D.); 11) a 2017 Schaefer Research Scholarship from the Vagelos College of Physicians and Surgeons (VP&S) of Columbia University (to P.D.); (12) NIH/NIDDK Grant K08-DK097181 (to Michael E. Rothenberg), NIH/NCI Grant R00-CA151673 (to Debashis Sahoo), NIH/NIGMS Grant R01-GM138385 (to D.S.), NIH/NIDCR Grant R01-DE028961 (to P.D.), and NIH/NINDS Grant R01-NS109858 (to Vincenzo A. Gennarino); (13) the Paul A. Marks Scholar Program of the College of Physicians and Surgeons (VP&S) of Columbia University (to V.A.G.): and (14) the Promotion and Mutual Aid Corporation for Private Schools of Japan (to Y.S.). Research reported in this publication was supported in part through NIH/NCI Cancer Center Support Grant P30-CA013696. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Funding Information: Yohei Shimono is co-inventor on a patent application owned by StanfordUniversity (US-20110021607), describing the use of miRNAs, including miR-200c, as biomarkers for the identification and therapeutic targeting of cancer stem cells. Yohei Shimono holds a financial relationship with a pharmaceutical company that might be considered relevant to this study: Quanticel Pharmaceuticals, now a fully owned subsidiary of Celgene and Bristol Myers Squibb (patent royalties, stock ownership). Piero Dalerba is co-inventor on patents and patent applications owned by the UniversityofMichigan (US-7723112, US-20140030786) and StanfordUniversity (US-9329170, US-09850483) and describing the use of EpCAM, CD44 and CD66a/CEACAM1 as bio-markers for the identification and differential purification of different subsets of colon epithelial cells. Piero Dalerba holds financial relationships with pharmaceutical and biotechnological companies that might be considered relevant to this study, including relationships with: Oncomed Pharmaceuticals, now a fully owned subsidiary of the Mereo BioPharma Group (patent royalties), Quanticel Pharmaceuticals, now a fully owned subsidiary of Celgene and Bristol Myers Squibb (patent royalties, stock ownership), Forty Seven Inc., now a fully owned subsidiary of Gilead Sciences Inc. (patent royalties, stock ownership), Amgen (stock ownership), Alexion Pharmaceuticals Inc. (employment of an immediate family member, stock ownership), AstraZeneca plc (stock ownership), Eli Lilly and Company (stock ownership), Merck & Co Inc. (stock ownership) and Pfizer Inc. (stock ownership). Piero Dalerba received a grant from BD Biosciences. Piero Dalerba received an honorarium from the Samsung Medical Center to give a scientific lecture. Michael E. Rothenberg is co-inventor on a patent application owned by StanfordUniversity (US-20130225435), describing the use of CEACAM1/CD66a as bio-marker for the identification and differential purification of different subsets of colon epithelial cells, and is currently an employee of Genentech, now a fully owned subsidiary of Roche Holding AG. Kaiqin Lao was an employee of Thermo Fisher Scientific, which commercializes some of the reagents used in this study for the analysis of miRNA expression levels, and currently serves as the chiefexecutiveofficer (CEO) of X Gen US. Shigeo Hisamori, Junko Mukohyama, Taichi Isobe, Luis E. Valencia Salazar, Xin Qian, Darius M. Johnston, Dalong Qian, Yoshihiro Kakeji, Vincenzo A. Gennarino and Debashis Sahoo disclose no conflicts of interest considered relevant to this study. Funding Information: The authors wish to thank Michael F. Clarke, MD (InstituteforStemCellBiologyandRegenerativeMedicine, StanfordUniversity) for his continuing mentorship and invaluable scientific insights. We thank Dr. Seetha V. Srinivasan (HerbertIrvingComprehensiveCancerCenter, ColumbiaUniversity) for editorial assistance during the preparation of this manuscript. We thank Yusuke Akama and Hiromi Yamazaki (TechnoProInc., FujitaHealthUniversity) and Hiroaki Sakai (FujitaHealthUniversity) for exceptional technical assistance. This work was supported by: (1) the JapanSocietyforthePromotionofScience (JSPS), through a ResearchFellowshipforYoungScientists (to Shigeo Hisamori), an OverseasResearchFellowship (to Junko Mukohyama) and the Grants-in-AidforScientificResearch (KAKENHI) program, with specific regard to grants 17K16555 (to J.M.), 15K14381 (to Yohei Shimono), 18K07231 (to Y.S.), 19K09106 (to Y.S.) and 21H02769 (to Y.S.); (2) the Japan-BelgiumResearchCooperativeProgram (to Y.S.); (3) a post-doctoral fellowship from the UeharaMemorialFoundation (to J.M.); (4) a post-doctoral scholarship from TheCellScienceResearchFoundation (to J.M.); (5) the EmpireStateInstitutionalTrainingProgram (DOH01-C30291GG-3450000) of the NewYorkStateStemCellScience (NYSTEM) agency (to J.M.); (6) a grant from the PrincessTakamatsuCancerResearchFund (to Y.S.); (7) a Grant from FujitaHealthUniversity (to Y.S.); (8) an ExtramuralCollaborativeResearchGrant from the CancerResearchInstitute of KanazawaUniversity (to Y.S.); (9) a Grant from the JapanAssociationforDevelopmentofCommunityMedicine (to T.I.); (10) a BDBiosciences 2011 StemCellResearchGrant (to Piero Dalerba); (11) a 2016 Runyon-RachleffInnovatorAward (DRR-44-16) from the DamonRunyonCancerResearchFoundation (to P.D.); 11) a 2017 SchaeferResearchScholarship from the Vagelos CollegeofPhysiciansandSurgeons (VP&S) of ColumbiaUniversity (to P.D.); (12) NIH/NIDDK Grant K08-DK097181 (to Michael E. Rothenberg), NIH/NCI Grant R00-CA151673 (to Debashis Sahoo), NIH/NIGMS Grant R01-GM138385 (to D.S.), NIH/NIDCR Grant R01-DE028961 (to P.D.), and NIH/NINDS Grant R01-NS109858 (to Vincenzo A. Gennarino); (13) the Paul A. Marks Scholar Program of the College of Physicians and Surgeons (VP&S) of Columbia University (to V.A.G.): and (14) the Promotion and Mutual Aid Corporation for Private Schools of Japan (to Y.S.). Research reported in this publication was supported in part through NIH/NCI Cancer Center Support Grant P30-CA013696. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2022, Japanese Society of Gastroenterology.
PY - 2022/6
Y1 - 2022/6
N2 - 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-derivedxenografts (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.
AB - 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-derivedxenografts (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.
UR - http://www.scopus.com/inward/record.url?scp=85125582038&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85125582038&partnerID=8YFLogxK
U2 - 10.1007/s00535-022-01865-9
DO - 10.1007/s00535-022-01865-9
M3 - Article
C2 - 35244768
AN - SCOPUS:85125582038
VL - 57
SP - 407
EP - 422
JO - Journal of Gastroenterology
JF - Journal of Gastroenterology
SN - 0944-1174
IS - 6
ER -