CRISPR-based gene disruption coupled to targeted integration of novel, high-avidity, natural T cell receptors to WT1 for acute leukemia

Newly identified WT1-specific TCRs can be employed to generate an army of engineered lymphocytes with redirected specificity towards WT1-expressing malignancies. T cells specific for the tumor antigen WT1 were expanded by sequential stimulation of peripheral blood mononuclear cells harvested from healthy donors with a pool of overlapping peptides spanning the full length of the protein. Longitudinal monitoring of the TCR alpha/beta clonotypes fluctuations enabled the retrieval of the anti-WT1 TCRs.