Cellular Barcoding to Monitor the Clonal Dynamics of ALL

Department of OncologyGregory Schwing

Gawad LabMentor: Dr Gawad, MD, PhD

St Jude8/7/15

Acute Lymphoblastic Leukemia

American Society of Hematology et al 2012

3000 children are diagnosed with ALL each year

• Approximately 600 cases of TELAML1 every year

• TELAML1 occurs in utero

• TELAML1 causes a halt in the development of B-cell progenitor cells

• 60,000 children are born with the TELAML1 mutation each year, a state we call Pre-Leukemic

ETV6-RUNX1

mRNA editingRetrovirus infection

2 functions:

(Harjes, Gross et al. 2009)

APOBEC

Hypothesized Model

Gawad et al 2014

• Unique heritable genetic tag• Frequency of barcode measures the fitness of a parent cell

Cellular Barcoding

GACTCTCTGAG

GACTCTCTGAG

GACTCTCTGAG

http://www.dreamstime.com/

Cellular Barcoding In Vivo Data

(Levy, Blundell et al. 2015)

GAG/POL REV/TAT ENV

Human Embryonic Kidney Cell

Transduction of Lenti-X

Dilution Factor

% G

FP E

xpre

ssio

n

• The barcoded vector was successfully produced in Lentivirus

• We determined the optimal concentration for viral transduction is the undiluted supernatant

Conclusion

• Analyze barcode diversity• Transduce the barcoded virus into the hematopoetic stem

cells, graft the HSC into mice, and monitor the barcodes.• Transduce APOBEC into the HSC coexpressed with ETV6-

RUNX1 to test our model

Future Directions

Acknowledgements