Open AccessIdentification, isolation, and characterization of human LGR5-positive colon adenoma cells
Author(s) -
Michael K. Dame,
Durga Attili,
Shan D. McClintock,
Priya H. Dedhia,
Peter Ouillette,
Olaf Hardt,
Alana M. Chin,
Xiang Xue,
Julie Laliberté,
Erica L. Katz,
Gina M. Newsome,
David R. Hill,
Alyssa J. Miller,
Yu-Hwai Tsai,
David Agorku,
Christopher Altheim,
Andreas Bosio,
B Simon,
Linda C. Samuelson,
Jay Stoerker,
Henry D. Appelman,
James Varani,
Max S. Wicha,
Dean E. Brenner,
Yatrik M. Shah,
Jason R. Spence,
Justin A. Colacino
Publication year - 2018
Publication title -
development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.754
H-Index - 325
eISSN - 1477-9129
pISSN - 0950-1991
DOI - 10.1242/dev.153049
Subject(s) - lgr5 , biology , organoid , stem cell , stem cell marker , wnt signaling pathway , carcinogenesis , cancer stem cell , microbiology and biotechnology , cancer research , gene , genetics , signal transduction
The intestine is maintained by stem cells located at the base of crypts and distinguished by the expression of LGR5. Genetically engineered mouse models have provided a wealth of information about intestinal stem cells, while less is known about human intestinal stem cells due to difficulty detecting and isolating these cells. We established an organoid repository from patient-derived adenomas, adenocarcinomas, and normal colon, which we analyzed for variants in 71 colorectal cancer (CRC) associated genes. Normal and neoplastic colon tissue organoids were analyzed by immunohistochemistry and fluorescent-activated cell sorting for LGR5. LGR5-positive cells were isolated from 4 adenoma organoid lines and were subjected to RNA-sequencing. We found that LGR5 expression in the epithelium and stroma was associated with tumor stage, and by integrating functional experiments with LGR5-sorted cell RNA-seq data from adenoma and normal organoids, we found correlations between LGR5 and CRC-specific genes, including DKK4 (dickkopf WNT signaling pathway inhibitor 4) and SMOC2 (SPARC related modular calcium binding 2). Collectively, this work provides resources, methods and new markers to isolate and study stem cells in human tissue homeostasis and carcinogenesis.