molecular barcoding in hps

Molecular Barcoding in HPS

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

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

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

Constructing the Vectors

TELAML

CONTROL

APOBEC3D

CONTROL

Model of Leukemogenesis

Single Cell Analysis96 Well C1 Chip

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

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

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