title: Use of methylation patterns to determine expansion of stem cell clones in human colon tissue.
abstract: OBJECTIVE
  It is a challenge to determine the dynamics of stem cells within human epithelial tissues such as colonic crypts. By tracking methylation patterns of nonexpressed genes, we have been able to determine how rapidly individual stem cells became dominant within a human colonic crypt. We also analyzed methylation patterns to study clonal expansion of entire crypts via crypt fission.
  
  
  METHODS
  Colonic mucosa was obtained from 9 patients who received surgery for colorectal cancer. The methylation patterns of Cardiac-specific homeobox, Myoblast determination protein 1, and Biglycan were examined within clonal cell populations, comprising either part of, or multiple adjacent, normal human colonic crypts. Clonality was demonstrated by following cytochrome c oxidase-deficient (CCO⁻) cells that shared an identical somatic point mutation in mitochondrial DNA.
  
  
  RESULTS
  Methylation pattern diversity among CCO⁻ clones that occupied only part of a crypt was proportional to clone size; this allowed us to determine rates of clonal expansion. Analysis indicated a slow rate of niche succession within the crypt. The 2 arms of bifurcating crypts had distinct methylation patterns, indicating that fission can disrupt epigenetic records of crypt ancestry. Adjacent clonal CCO⁻ crypts usually had methylation patterns as dissimilar to one another as methylation patterns of 2 unrelated crypts. Mathematical models indicated that stem cell dynamics and epigenetic drift could account for observed dissimilarities in methylation patterns.
  
  
  CONCLUSIONS
  Methylation patterns can be analyzed to determine the rates of recent clonal expansion of stem cells, but determination of clonality over many decades is restricted by epigenetic drift. We developed a technique to follow changes in intestinal stem cell dynamics in human epithelial tissues that might be used to study premalignant disease.
authors.affiliation: Histopathology Laboratory, Cancer Research UK London Research Institute, London, UK. trevor.graham@cancer.org.uk
authors.forename: Trevor A
authors.initials: TA
authors.lastname: Graham
authors.forename: Adam
authors.initials: A
authors.lastname: Humphries
authors.forename: Theodore
authors.initials: T
authors.lastname: Sanders
authors.forename: Manuel
authors.initials: M
authors.lastname: Rodriguez-Justo
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Tadrous
authors.forename: Sean L
authors.initials: SL
authors.lastname: Preston
authors.forename: Marco R
authors.initials: MR
authors.lastname: Novelli
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Leedham
authors.forename: Stuart A C
authors.initials: SA
authors.lastname: McDonald
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Wright
mesh_terms: D053687:Adult Stem Cells / Q000502:physiology*; D058578:Biglycan / Q000235:genetics; D019070:Cell Lineage / Q000502:physiology; D002999:Clone Cells / Q000166:cytology / Q000502:physiology; D003106:Colon / Q000166:cytology* / Q000502:physiology*; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D019175:DNA Methylation / Q000502:physiology*; D004272:DNA, Mitochondrial / Q000235:genetics; D003576:Electron Transport Complex IV / Q000378:metabolism; D044127:Epigenesis, Genetic / Q000502:physiology; D040961:Genetic Drift; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000502:physiology; D017570:MyoD Protein / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011230:Precancerous Conditions / Q000235:genetics* / Q000473:pathology
chemical_list: C546884:BGN protein, human; D058578:Biglycan; D004272:DNA, Mitochondrial; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D003576:Electron Transport Complex IV