Modulation of Target Antigen Density Improves CAR T-cell Functionality and Persistence.

Abstract

Chimeric antigen receptor T-cell (CART) therapy targeting CD22 induces remission in 70% of patients with relapsed/refractory acute lymphoblastic leukemia (ALL). However, the majority of post-CD22 CART remissions are short and associated with reduction in CD22 expression. We evaluate the implications of low antigen density on the activity of CD22 CART and propose mechanisms to overcome antigen escape. Using ALL cell lines with variable CD22 expression, we evaluate the cytokine profile, cytotoxicity, and <i>in vivo</i> CART functionality in the setting of low CD22 expression. We develop a high-affinity CD22 chimeric antigen receptor (CAR) as an approach to improve CAR sensitivity. We also assess Bryostatin1, a therapeutically relevant agent, to upregulate CD22 and improve CAR functionality. We demonstrate that low CD22 expression negatively impacts <i>in vitro</i> and <i>in vivo</i> CD22 CART functionality and impairs <i>in vivo</i> CART persistence. Moreover, low antigen expression on leukemic cells increases naïve phenotype of persisting CART. Increasing CAR affinity does not improve response to low-antigen leukemia. Bryostatin1 upregulates CD22 on leukemia and lymphoma cell lines for 1 week following single-dose exposure, and improves CART functionality and <i>in vivo</i> persistence. While Bryostatin1 attenuates IFNγ production by CART, overall <i>in vitro</i> and <i>in vivo</i> CART cytotoxicity is not adversely affected. Finally, administration of Bryostain1 with CD22 CAR results in longer duration of <i>in vivo</i> response. We demonstrate that target antigen modulation is a promising strategy to improve CD22 CAR efficacy and remission durability in patients with leukemia and lymphoma.<i>See related commentary by Guedan and Delgado, p. 5188</i>.