Enhanced Tumor Trafficking of GD2 Chimeric Antigen Receptor T Cells by Expression of the Chemokine Receptor CCR2b

Abstract

For adoptive T cell therapy to be effective against solid tumors, tumor-specific T cells must be able to migrate to the tumor site. One requirement for efficient migration is that the effector cells express chemokine receptors that match the chemokines produced either by tumor or tumorassociated cells. In this study, we investigated whether the tumor trafficking of activated T cells (ATCs) bearing a chimeric antigen receptor specific for the tumor antigen GD2 (GD2-CAR) could be enhanced by forced co-expression of the chemokine receptor CCR2b, since this receptor directs migration towards CCL2, a chemokine produced by many tumors, including neuroblastoma. Neuroblastoma cell lines (SK-N-SH and SK-N-AS) and primary tumor cells isolated from six patients all secreted high levels of CCL2, but GD2-CAR transduced ATCs lacked expression of CCR2 (<5%) and migrated poorly to recombinant CCL2 or tumor supernatants. Following retroviral transduction, however, ATCs expressed high levels of CCR2b (>60%) and migrated well in vitro. We expressed firefly luciferase in CCR2b-expressing ATCs and observed improved homing (>10-fold) to CCL2-secreting neuroblastoma compared to CCR2 negative ATCs. As a result, ATCs co-modified with both CCR2b and GD2-CAR had greater anti-tumor activity in vivo.

DAP12-Based Activating Chimeric Antigen Receptor for NK Cell Tumor Immunotherapy

“…In the future it might therefore be worthwhile to genetically modify NK cells with chemokine receptors to enhance selective tumor infiltration. That this kind of engrafted chemotaxis is beneficial was previously demonstrated by CXCR2-and CCR2-modified T cells, which showed improved tumor infiltrations and eradications in tumor xenograft models (72,73).…”

Synthetic control of mammalian-cell motility by engineering chemotaxis to an orthogonal bioinert chemical signal

“…Other groups have described interesting approaches, including the chemical or enzymatic modification of the cell surface with specific adhesion molecules (45), materials engineering of artificial scaffolds and tunable matrices to direct cell adhesion and migration (46), expression or direct injection of natural homing ligands such as chemokines into sites where increased cell migration is desired (47,48), and the expression in therapeutic cells of natural receptors such as chemokine receptors whose ligands are up-regulated in inflammation or cancer (8,7,16,17). These strategies rely on naturally existing homing receptors and ligands, and they are powerful because they tap into cells' native migration axes.…”

“…The use of cells for the treatment of a growing array of diseases including cancer, autoimmunity, and chronic wounds is currently being explored (1)(2)(3)(4)(5)(6). The appropriate and efficient localization of therapeutic cells to sites of disease has been identified as an important factor for successful cell-based therapy (7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). However, preclinical studies and clinical trials to date have shown that the homing to sites of disease of many cell types commonly used as therapeutics is frequently impaired or limited, especially after ex vivo expansion of cells in culture (7,12,18,19).…”

Genetic modification of human T lymphocytes for the treatment of hematologic malignancies

“…For example, specific chemokine receptors can be ectopically expressed by T cells to improve their homing along the chemokine gradients produced by tumor cells. This approach is effective in pre-clinical models of melanoma, Hodgkin's lymphoma, and neuroblastoma in which overexpression of CXCR2, 77 CCR4 78 or CCR2b 79 by T cells enhanced their trafficking (and subsequent antitumor effects) along CXCL1, TARC and CCL2 gradients, respectively. Genetic modifications have also been exploited to counter inhibitory molecules present in the tumor microenvironment.…”