molecular barcoding in hps
TRANSCRIPT
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Molecular Barcoding in HPS
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Leukemia is Heterogeneous
• Stage of development, mutations, and clonal fitness• The cancer consists of a combination of index clones and
related clones• Relapse is often from the emergence of a related clone
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ETV6-RUN1X
• Occurs in utero• A fusion of the DBD of RUNX1 and the full ETV6 transcription
factor• Halts development of lymphoid cells causing a buildup of b-cell
progenitors• Initiates a pre-leukemic condition
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APOBEC
• Apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like cytosine deaminase
• C to U mutations• Active during RNA editing and retrovirus infections• Body wide mutagenesis
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Constructing the Vectors
TELAML
CONTROL
APOBEC3D
CONTROL
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Model of Leukemogenesis
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Single Cell Analysis96 Well C1 Chip
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Data AnalysisWe could roughly detect a 1% prevalence clone with 200 cells, 2% with 75 cells, and 4% with 50 cells. Thus, on average, we would roughly need to identify at least 2–3 distinct cells from the same clone to accurately detect that population
The barcodes will be used to separate the clones and monitor the fitness of each clones
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Conclusion
• The heterogeneity of leukemia requires constant monitoring of the tumor structure
• By modeling Leukemogenesis we can predict what clones will have the highest fitness in the clinic
• We accomplish this by isolating the barcodes with the highest frequency and sequencing them
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Acknowledgements
• Dr Charles Gawad, M.D., Ph D• Dr. Veronica Gonzalez-Peña, Ph D• Mr. Shelby Lane, B.S.• Dr. Gronemeyer• Mr. James Marmion• My family, friends, and fellow POE’s