All cancers were once normal cells. They became cancerous through the chance acquisition of particular somatic mutations that gave them a selective advantage over their neighbours. Thus, the mutations that initiate cancer occur in normal cells, and the normal clonal dynamics of the tissue determine a mutant cell's ability to establish a malignant clone; yet these remain poorly understood in humans. One tissue was selected for the exploration of each of these two facets of somatic evolution: blood for clonal dynamics; colon for mutational processes. Blood presents an opportunity to study normal human clonal dynamics, as clones mix spatially and longitudinal samples can be taken. We isolated 140 single haematopoietic stem and progenitor cells from a healthy 59 year-old and grew them in vitro into colonies that were whole genome sequenced. Population genetics approaches were applied to this dataset, allowing us to elucidate for the first time the number of active haematopoietic stem cells, the rate at which clones grow and shrink, and the cellular output of stem cell clones. Colonic epithelium is organised into crypts, at the base of which sit a small number of stem cells. All cells in a crypt ultimately share an ancestor in one stem cell that existed recently, and consequently share the mutations that were present in this ancestor. We exploited this natural clonal unit, isolating single colonic crypts through laser capture microdissection. 570 colonic crypts from 42 individuals were whole genome sequenced. We describe the burden and pattern of somatic mutations in these genomes and their variability across and within different people, identifying some mutational processes that are ubiquitous and others that are sporadic. Targeted sequencing of an additional 1,500 crypts allowed us to quantify the frequency of driver mutations in normal human colon. Together, these two studies inform on the somatic evolution of normal tissues, describing new biology in human tissue homeostasis and providing a window into the processes that govern cancer incidence.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:767860 |
| Date | January 2019 |
| Creators | Lee-Six, Henry |
| Contributors | Campbell, Peter ; Stratton, Michael |
| Publisher | University of Cambridge |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://www.repository.cam.ac.uk/handle/1810/289819 |
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