title: Transcriptional Regulator CNOT3 Defines an Aggressive Colorectal Cancer Subtype.
abstract: Cancer cells exhibit dramatic alterations of chromatin organization at cis-regulatory elements, but the molecular basis, extent, and impact of these alterations are still being unraveled. Here, we identify extensive genome-wide modification of sites bearing the active histone mark H3K4me2 in primary human colorectal cancers, as compared with corresponding benign precursor adenomas. Modification of certain colorectal cancer sites highlighted the activity of the transcription factor CNOT3, which is known to control self-renewal of embryonic stem cells (ESC). In primary colorectal cancer cells, we observed a scattered pattern of CNOT3 expression, as might be expected for a tumor-initiating cell marker. Colorectal cancer cells exhibited nuclear and cytoplasmic expression of CNOT3, suggesting possible roles in both transcription and mRNA stability. We found that CNOT3 was bound primarily to genes whose expression was affected by CNOT3 loss, and also at sites modulated in certain types of colorectal cancers. These target genes were implicated in ESC and cancer self-renewal and fell into two distinct groups: those dependent on CNOT3 and MYC for optimal transcription and those repressed by CNOT3 binding and promoter hypermethylation. Silencing CNOT3 in colorectal cancer cells resulted in replication arrest. In clinical specimens, early-stage tumors that included >5% CNOT3+cells exhibited a correlation to worse clinical outcomes compared with tumors with little to no CNOT3 expression. Together, our findings implicate CNOT3 in the coordination of colonic epithelial cell self-renewal, suggesting this factor as a new biomarker for molecular and prognostic classification of early-stage colorectal cancer. Cancer Res; 77(3); 766-79. ©2016 AACR.
authors.affiliation: Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts.|Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.|Department of Medical Oncology, Hospital La Paz Institute for Health Research CIBERONC, Madrid, Spain.
authors.forename: Paloma
authors.initials: P
authors.lastname: Cejas
authors.affiliation: Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts.|Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.
authors.forename: Alessia
authors.initials: A
authors.lastname: Cavazza
authors.affiliation: Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts.
authors.forename: C N
authors.initials: CN
authors.lastname: Yandava
authors.affiliation: Department of Medical Oncology, Hospital La Paz Institute for Health Research CIBERONC, Madrid, Spain.
authors.forename: Victor
authors.initials: V
authors.lastname: Moreno
authors.affiliation: Pathology Institute, Ludwig-Maximilians-Universitat, Munich, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Horst
authors.affiliation: Department of Medical Oncology, Hospital La Paz Institute for Health Research CIBERONC, Madrid, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Moreno-Rubio
authors.affiliation: Department of Pathology, Hospital La Paz Institute for Health Research, Madrid, Spain.
authors.forename: Emilio
authors.initials: E
authors.lastname: Burgos
authors.affiliation: Department of Medical Oncology, Hospital La Paz Institute for Health Research CIBERONC, Madrid, Spain.|Department of Pathology, Hospital La Paz Institute for Health Research, Madrid, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Mendiola
authors.affiliation: Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts.
authors.forename: Len
authors.initials: L
authors.lastname: Taing
authors.affiliation: Center for Gastrointestinal Cancer Research, Baylor University Medical Center, Dallas, Texas.
authors.forename: Ajay
authors.initials: A
authors.lastname: Goel
authors.affiliation: Department of Medical Oncology, Hospital La Paz Institute for Health Research CIBERONC, Madrid, Spain.
authors.forename: Jaime
authors.initials: J
authors.lastname: Feliu
authors.affiliation: Department of Medical Oncology and Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Boston, Massachusetts. ramesh_shivdasani@dfci.harvard.edu.|Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts.
authors.forename: Ramesh A
authors.initials: RA
authors.lastname: Shivdasani
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis; D047369:Chromatin Immunoprecipitation; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000401:mortality / Q000473:pathology*; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000473:pathology; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D051379:Mice; D016688:Mice, Inbred NOD; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046888:Tissue Array Analysis; D014157:Transcription Factors / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D014408:Biomarkers, Tumor; C466622:CNOT3 protein, human; D014157:Transcription Factors