SLIDE 1
Krzysztof Skiba 09.12 2015 Wrocaw What is SCS? History of SCS - - PowerPoint PPT Presentation
Krzysztof Skiba 09.12 2015 Wrocaw What is SCS? History of SCS - - PowerPoint PPT Presentation
Krzysztof Skiba 09.12 2015 Wrocaw What is SCS? History of SCS Clonal diversity and evolution in primary tumors Circulating tumor cells and metastatic dissemination Delineating complex chromosome rearrangements in single
SLIDE 2
SLIDE 3
Single cell sequencing NGS on single cells Huge impact on cancer research
Differences to NGS?
Single cell extraction WGA (whole genome amplification) before
NGS (genomic input in ng)
Looped amplicons
SLIDE 4
SLIDE 5
SLIDE 6
Intratumor heterogeneity (phylogenetic lineages
for cells in tumor mass)
Clonal evolution:
- by SCS
- mostly CNVs in early tumor cells development
- SNPs accumulation over time
Single cells share common founder mutations,
suggesting evolution from a common origin
SLIDE 7
Cells that circulate in blood RARE (1 cel in 1 000 000) Carry ~60% SNPs from primary and
metastatic tumors (prostate, lung, breast)
Blood markers for tumors? (non invasive) CTCs express their own extracellular matrix
proteins in the blood
SLIDE 8
Tumor evolution = chromosome structural
rearrangments = gene dosage effects
Evolution dynamics SCS represents cells at differnet evolution
time
CNA – copy number aberrations, early in
tumor evolution
Chromothripsis – within single chromosome
SLIDE 9
Initial response to therapy, but eventually
resistance to treatment
Epithelial-to-mesenchymal transition (EMT) Resistance clones preexisting (adaptive) Induced new mutations (acquired) SCS: Androgen gene amplification in CTCs
(prostate cancer = adaptive resistance)
SLIDE 10
SLIDE 11
Human cancers = higher mutation rate ? Estimated mutation rate: 210 x normal cells SCS: 2.5 nucleotide error per cell division
marginally higher then normal
SCS: breast cancer – 13.3 x normal SCS limited to few patients – need more
research
SLIDE 12
Genetically engineered mice (GEM) Xenografts SCS: transplantation of breast cancer from
human to mouse1 -> mouse2 ->… mouseN. Selection of tumor cells and high mutation rate.
RNA-SCS: pancreatic cancer, CTCs and
fluorescent markers.
SLIDE 13
Phylogenetic trees -> therapeutic targets CTCs and cancer cells in bodily fluids -> early
noninvasive detection
SCS advantage -> very small clinical samples
SLIDE 14
Drop-seq (uses nanoliter droplets and
barcoded beads to capture single cells and performWTA, enabling RNA-SCS on 10,000– 100,000 cells in a single run )
https://vimeo.com/128484564
SLIDE 15
SLIDE 16
SCS improoved understanding of
decelopment , heterogeneity, metastasis, invasion of tumor cells.
SCS for gene expression analysis. SCS as a tool for connecting genotypes with
phenotypes.
SLIDE 17
The first five years of single-cell cancer genomics and beyond Nicholas E. Navin, doi:10.1101/gr.191098.115Genome Res. 2015. 25:1499-1507