Poster #019
Fluorescent Protein Barcoding Reveals Heterogeneity in Leukemia Cells
Mentors: Katsuyuki Nishi, Ph.D.; Ravi Majeti, M.D., Ph.D.
Leukemia is the most common cancer among children and teens. Studies have shown that leukemia stem cells (LSCs) are typically composed of multiple subclones with distinct characteristics. This heterogeneity poses a significant therapeutic challenge as varied responses contribute to relapse and resistance. Conventional methods for analyzing heterogeneity involve DNA/RNA extractions, which kills cells and restricts further analysis. To address this, we hypothesized that using fluorescent protein barcoding via gene-editing technology would distinguish subpopulations within a human leukemic cell line (TF-1 cells). We introduced the multi-fluorescence Zebrabow system by infecting TF-1 cells with lentivirus, followed by electroporation of a plasmid encoding Cre protein, which induces expression of different fluorescent proteins. Using fluorescent activated cell sorting of live cells, we found that each LSC subclone expressed one of three colors, allowing us to identify and trace distinct subpopulations. This approach may enable better investigations into stem cell heterogeneity and therapy resistance.