Deterministic fusion and single cell sequencing of heterokaryons
Cellular fusion is a fundamental process that regulates a wide variety of essential functions such as fertilization, musculoskeletal tissue development, immune response and antibody production, tumorigenesis and neuronal signaling. The fusion of two cells occurs through cell-cell contact, cell wall merging and remodeling followed by mixing of cytoplasmic material. Understanding fusion mechanisms has been hampered by several challenges that prevent quantitative insights such as heterogeneity. To answer fundamental and applied questions about cellular fusion that drive tumor heterogeneity, we propose to develop a program that fuses cancerous cells with healthy cells of different types and read out the consequences of fusion using single cell sequencing. To accomplish this task, we will utilize bi-species heterokaryons (mouse and human) as test cellular systems from different cancer cell lines and evaluate their tumor forming potential by transplantation into mice. Fusing diseased cells with healthy cells will permit, for the first time, insights into safeguards or expression of proteins that prevent fusion of dysregulated cells and how fusion can drive acquisition of deleterious traits such as increased metastatic behavior.