Adoptive Transfer of Chimeric Antigen Receptor Re-directed Cytolytic T Lymphocyte Clones in Patients with Neuroblastoma

Abstract

Metastatic neuroblastoma is a poor-prognosis malignancy arising during childhood that overexpresses the L1-cell adhesion molecule (CD171). We have previously described a tumor L1-cell adhesion molecule-specific, single chain antibody-derived, chimeric antigen receptor designated CE7R for re-directing the antigen-specific effector functioning of cytolytic T lymphocytes. Here, we report on the feasibility of isolating, and the safety of infusing, autologous CD8(+) cytolytic T lymphocyte clones co-expressing CE7R and the selection-suicide expression enzyme HyTK in children with recurrent/refractory neuroblastoma. The cytolytic T lymphocyte products were derived from peripheral blood mononuclear cells that were subjected to polyclonal activation, plasmid vector electrotransfer, limiting dilution hygromycin selection, and expansion to numbers sufficient for adoptive transfer. In total, 12 infusions (nine at 10(8) cells/m(2), three at 10(9) cells/m(2)) were administered to six patients. No overt toxicities to tissues known to express L1-cell adhesion molecule (e.g., central nervous system, adrenal medulla, and sympathetic ganglia) were observed. The persistence of cytolytic T lymphocyte clones in the circulation, measured by vector-specific quantitative polymerase chain reaction, was short (1-7 days) in patients with bulky disease, but significantly longer (42 days) in a patient with a limited disease burden. This first-in-humans pilot study sets the stage for clinical trials employing adoptive transfer in the context of minimal residual disease.

Coexpressed Catalase Protects Chimeric Antigen Receptor–Redirected T Cells as well as Bystander Cells from Oxidative Stress–Induced Loss of Antitumor Activity

“…Recently, transfer of such second generation CAR T cells targeting CD19 + B cell lymphoid leukemia has shown encouraging clinical results in treating patients with bulky tumors (7)(8)(9)(10). Although these results are galvanizing the field of adoptive cell therapy, clinical trials focusing on solid tumors have seen less success (11)(12)(13). The challenge for T cell-based therapies of solid tumors lies in that T cells, in addition to reaching their targets, are required to survive and function within the unfavorable tumor microenvironment.…”

Infusing CD19-Directed T Cells to Augment Disease Control in Patients Undergoing Autologous Hematopoietic Stem-Cell Transplantation for Advanced B-Lymphoid Malignancies

“…To overcome immune tolerance to tumor-associated antigens (TAAs) investigators have developed and implemented immunotherapies infusing T cells genetically modified to express a chimeric antigen receptor (CAR) (Paulos et al, 2008;Jena et al, 2010). CARs are typically engineered to provide the genetically modified T cell with the specificity of a murine monoclonal antibody and, on binding a cell surface TAA, can specifically activate the T cell for proliferation, cytokine production, and cytolysis (Eshhar et al, 1993;Brentjens et al, 2003;Cooper et al, 2003;Park et al, 2007;Brenner and Heslop, 2010;Porter et al, 2011). CARs are ''universal'' in that they bind TAAs independent of major histocompatibility complex (MHC), and thus one receptor construct can be used to treat a population of patients with the same TAA + tumors.…”

The Optimal Antigen Response of Chimeric Antigen Receptors Harboring the CD3ζ Transmembrane Domain Is Dependent upon Incorporation of the Receptor into the Endogenous TCR/CD3 Complex

“…R edirecting T cell specificity through the expression of artificial Ag receptors is currently the source of widespread research and testing in a number of early phase clinical trials worldwide (1,2). Two basic approaches have been explored to date: first, the expression of a and b TCR chains in the T cell, thereby driving the formation of new TCR complexes with the desired Ag specificity in the gene-modified T cell (3,4); and second, the expression of receptors bearing T cell signaling receptors fused to extracellular domains that bind to defined target Ags (5).…”