Molecular engineering to enhance car-t mediated tumor immunotherapy

Abstract

The present disclosure provides a chimeric antigen receptor (CAR) immunotherapy for treating a hematologic cancer comprising (a) genetically modifying a population of immune effector cells comprising T cell receptors (TCRs) to stably express at least one CAR, wherein the ectodomain of the CAR specifically binds a cancer antigen; (b) expanding the population of CAR-containing immune effector cells in the presence of one or more cytokines in vitro to achieve a therapeutic dose, wherein the population of CAR-containing immune effector cells exhibits a nonexhausted memory T cell phenotype; and (c) infusing eligible subjects with the CAR-containing immune effector memory T cell phenotype population of cells as needed until the hematologic cancer in the body is destroyed. The intracellular signal transduction domain of the CAR includes 4-1BB, a CD3ζ T cell activation chain and a genetically engineered TLE3, which can be ectopically expressed. The immune effector cell population engineered to express the CAR comprising the engineered TLE3 construct by lentiviral transduction is a CD3+ T cell population or an NK cell. Expression of the engineered TLE3 construct in stimulated CAR-containing immune effector cells in vitro and in vivo xenograft preclinical models promotes activation of genes that promote anti-tumor activity and also represses genes involved in excessive effector function, exhaustion or both.