Cell-Based Models of 'Cytokine Release Syndrome' Endorse CD40L and Granulocyte-Macrophage Colony-Stimulating Factor Knockout in Chimeric Antigen Receptor T Cells as Mitigation Strategy

• Published in: Cells (Basel, Switzerland) Vol. 12; no. 21; p. 2581
• Main Authors: Dibas, Ala, Rhiel, Manuel, Patel, Vidisha Bhavesh, Andrieux, Geoffroy, Boerries, Melanie, Cornu, Tatjana I, Alzubi, Jamal, Cathomen, Toni
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.11.2023
• Subjects: Analysis; Animal models; Animals; Antibodies; Antigens; Blood & organ donations; CD19 antigen; CD19-CAR T cells; CD40 Ligand; CD40L; CD40L protein; Cell activation; Cell culture; Cell therapy; Cells; Chimeric antigen receptors; Colony-stimulating factor; CRISPR; CRISPR-Cas9; CRS; Cytokine Release Syndrome; Cytokines; Cytolytic activity; Drug dosages; Genomes; GM-CSF; Granulocyte-macrophage colony-stimulating factor; Granulocyte-Macrophage Colony-Stimulating Factor - genetics; Granulocytes; Humans; Immunity; Inflammation; Interleukin 6; knockout; Leukemia; Lymphocytes; Lymphocytes B; Lymphocytes T; Macrophages; Malignancy; Mice; Mice, Knockout; Monocytes; Receptors, Chimeric Antigen - genetics; Steroids; T-Lymphocytes; Testing; Tumor antigens; Tumor necrosis factor-TNF; Germany; St Louis Missouri; United States > US; CD40L; knockout; CRS; GM-CSF; CRISPR-Cas9; CD19-CAR T cells
• DOI: 10.3390/cells12212581
• ISSN: 2073-4409

While chimeric antigen receptor (CAR) T cell therapy has shown promising outcomes among patients with hematologic malignancies, it has also been associated with undesirable side-effects such as cytokine release syndrome (CRS). CRS is triggered by CAR T-cell-based activation of monocytes, which are stimulated via the CD40L-CD40R axis or via uptake of GM-CSF to secrete proinflammatory cytokines. Mouse models have been used to model CRS, but working with them is labor-intensive and they are not amenable to screening approaches. To overcome this challenge, we established two simple cell-based CRS in vitro models that entail the co-culturing of leukemic B cells with CD19-targeting CAR T cells and primary monocytes from the same donor. Upon antigen encounter, CAR T cells upregulated CD40L and released GM-CSF which in turn stimulated the monocytes to secrete IL-6. To endorse these models, we demonstrated that neutralizing antibodies or genetic disruption of the and/or loci in CAR T cells using CRISPR-Cas technology significantly reduced IL-6 secretion by bystander monocytes without affecting the cytolytic activity of the engineered lymphocytes in vitro. Overall, our cell-based models were able to recapitulate CRS in vitro, allowing us to validate mitigation strategies based on antibodies or genome editing.