Generation of Tumor-Reactive T Cells by Co-culture of Peripheral Blood Lymphocytes and Tumor Organoids

Abstract

Cancer immunotherapies have shown substantial clinical activity for a subset of patients with epithelial cancers. Still, technological platforms to study cancer T-cell interactions for individual patients and understand determinants of responsiveness are presently lacking. Here, we establish and validate a platform to induce and analyze tumor-specific T cell responses to epithelial cancers in a personalized manner. We demonstrate that co-cultures of autologous tumor organoids and peripheral blood lymphocytes can be used to enrich tumor-reactive T cells from peripheral blood of patients with mismatch repair-deficient colorectal cancer and non-small-cell lung cancer. Furthermore, we demonstrate that these T cells can be used to assess the efficiency of killing of matched tumor organoids. This platform provides an unbiased strategy for the isolation of tumor-reactive T cells and provides a means by which to assess the sensitivity of tumor cells to T cell-mediated attack at the level of the individual patient.

3D model for CAR ‐mediated cytotoxicity using patient‐derived colorectal cancer organoids

“…Testing in patient-matched normal and tumor organoids may greatly assist to further adapt these strategies. As a demonstration of the great potential of organoid systems for the development of cancer immunotherapies, tumor-reactive T cells have recently been expanded from peripheral blood using cocultures of matched CRC organoids (Dijkstra et al, 2018). While this strategy was only successful in a fraction of HLA-I-positive tumors and only in MSI patients, future CAR-T approaches might use organoids to selectively expand tumor-reactive cells for a larger fraction of patients.…”

“…Addition of stromal cells further allows to reconstruct the tissue microenvironment and to study immuno-epithelial crosstalk (Farin et al, 2014;Rogoz et al, 2015;Nozaki et al, 2016;Noel et al, 2017). Moreover, tumor-reactive T cells can be selectively expanded in co-culture with tumor organoids from MSI patients (Dijkstra et al, 2018).…”

Reflection of neuroblastoma intratumor heterogeneity in the new OHC‐NB1 disease model

“…A shortcoming of classical cancer organoid cultures is the lack of stroma, blood vessels and immune cells. More recently, coculture systems have incorporated cancer‐associated fibroblasts into pancreatic cancer organoids and matched tumor‐derived peripheral blood lymphocytes into colon and non‐small cell lung cancer organoids . Organoid culture systems are, however, more costly than 2D cultures, due to the need for extracellular matrix.…”

“…More recently, coculture systems have incorporated cancerassociated fibroblasts into pancreatic cancer organoids and matched tumor-derived peripheral blood lymphocytes into colon and non-small cell lung cancer organoids. 46,47 Organoid culture systems are, however, more costly than 2D cultures, due to the need for extracellular matrix. Here, we show that establishing OHC-NB1 neuroblastoma spheroids, as defined by spherical aggregates of tumor cells that originate from a tiny clinical bone marrow sample containing metastatic neuroblastoma cells, is technically feasible.…”

The tumour microenvironment shapes dendritic cell plasticity in a human organotypic melanoma culture

“…However, these matrices, while ensuring the required tissue stiffness, may otherwise act as a source of foreign antigens 39 . Thus, de-cellularized human tissue like the one here employed, or synthetic matrices 40 potentially represents better alternatives for establishing an immunocompetent TME.…”