Fisher, Jonathan2019-12-182019-052019-06-282019Fisher, Jonathan. 2019. Stem Cell Hierarichies within Tumor Genetic Subclones. Doctoral dissertation, Harvard Medical School.http://nrs.harvard.edu/urn-3:HUL.InstRepos:42063324Intratumoral heterogeneity has been appreciated at the genetic and transcriptional levels, yet it remains unclear how they relate to one another. My thesis project applies a novel qPCR technique to combine single-cell RNA-seq with same-cell cDNA genotyping at single-nucleotide resolution. Using whole-exome sequencing in a human oligodendroglioma tumor, we identified a mutation in CIC that is present in only a subset of cancer cells. A previous analysis of single-cell RNA-seq in this tumor showed a substantial minority of cancer cells that display a stemness phenotype distinct from differentiated lineage phenotypes previously known in low-grade glioma. Results of parallel qPCR genotyping demonstrate that both CIC-wild-type and CIC-mutant cells are found across the spectrum of stem-differentiated phenotypes. This suggests that the stemness phenotype exists independently of mutation status and that it is instead more likely epigenetically driven.application/pdfensingle-cell RNA-seqoligodendrogliomagliomatumor heterogeneitycancer stem cellsThesis or Dissertation2019-12-18