Selective Expansion of Chimeric Antigen Receptor-targeted T-cells with Potent Effector Function using Interleukin-4

• DOI: 10.1074/jbc.m110.127951
• Publication Date: 2010-06-20T23:48:33Z
• Authors: Scott Wilkie; Sophie E. Burbridge; Laura Chiapero-St...; Ana C.P. Pereira; Siobhán Cleary; Sjoukje J.C. van ...; James F. Spicer; David M. Davies; John Maher
• Keywords: Interleukin 3

Polyclonal T-cells can be directed against cancer using transmembrane fusion molecules known as chimeric antigen receptors (CARs). Although preclinical studies have provided encouragement, pioneering clinical trials CAR-based immunotherapy been disappointing. Key obstacles are the need for robust expansion ex vivo followed by sustained survival of infused in patients. To address this, we developed a system to achieve selective proliferation CAR(+) IL-4, cytokine with several pathophysiologic and therapeutic links cancer. A receptor (4alphabeta) was engineered IL-4 alpha (IL-4Ralpha) ectodomain beta(c) subunit, used IL-2 IL-15. Addition that express 4alphabeta resulted STAT3/STAT5/ERK phosphorylation exponential proliferation, mimicking actions IL-2. Using receptor-selective muteins, partnering gamma(c) implicated signal delivery. Next, human were co-express CAR specific tumor-associated MUC1. These exhibited an unprecedented capacity elicit repeated destruction MUC1-expressing tumor cultures expanded through logs vitro. Despite prolonged culture retained specificity target antigen, type 1 polarity, dependence. Similar findings observed CARs two additional targets, demonstrating generality application. Furthermore, this allows rapid enrichment from small blood volumes, under GMP-compliant conditions. Together, these provide proof principle development IL-4-enhanced T-cell