Granulocyte–macrophage colony-stimulating factor inactivation in CAR T-cells prevents monocyte-dependent release of key cytokine release syndrome mediators

Abstract

Edited by Peter Cresswell Chimeric antigen receptor T-cell (CAR T-cell) therapy has been shown to be clinically effective for managing a variety of hematological cancers. However, CAR T-cell therapy is associated with multiple adverse effects, including neurotoxicity and cytokine release syndrome (CRS). CRS arises from massive cytokine secretion and can be life-threatening, but it is typically managed with an anti-IL-6Ra mAb or glucocorticoid administration. However, these treatments add to a patient's medication burden and address only the CRS symptoms. Therefore, alternative strategies that can prevent CRS and neurotoxicity associated with CAR T-cell treatment are urgently needed. Here, we explored a therapeutic route aimed at preventing CRS rather than limiting its consequences. Using a cytokine-profiling assay, we show that granulocyte-macrophage colony-stimulating factor (GMCSF) is a key CRS-promoting protein. Through a combination of in vitro experiments and gene-editing technology, we further demonstrate that antibody-mediated neutralization or TALEN-mediated genetic inactivation of GMCSF in CAR T-cells drastically decreases available GMCSF and abolishes macrophage-dependent secretion of CRS biomarkers, including monocyte chemoattractant protein 1 (MCP-1), interleukin (IL) 6, and IL-8. Of note, we also found that the genetic inactivation of GMCSF does not impair the antitumor function or proliferative capacity of CAR T-cells in vitro. We conclude that it is possible to prevent CRS by using "all-in-one" GMCSF-knockout CAR T-cells. This approach may eliminate the need for anti-CRS treatment and may improve the overall safety of CAR T-cell therapies for cancer patients.

Glial injury in neurotoxicity after pediatric CD19‐directed chimeric antigen receptor T cell therapy

“…10,[30][31][32] These cytokines have also been implicated in a number of other CNS diseases 33 and are known to activate the endothelial cells at the BBB, 9,34,35 supporting the theory that ICANS is initiated by a systemic cytokine surge. Key inflammatory cytokines showed similar elevations in the CSF and serum, suggesting that local cytokine production in the CNS is not a prominent feature of neurotoxicity.…”

“…Evidence is emerging that T cells may activate monocytes through granulocyte macrophage colony-stimulating factor and that the monocytes are an important source of proinflammatory cytokines. 10,[30][31][32] These cytokines have also been implicated in a number of other CNS diseases 33 and are known to activate the endothelial cells at the BBB, 9,34,35 supporting the theory that ICANS is initiated by a systemic cytokine surge.…”

De novodesigned transmembrane domains tune engineered receptor functions

“…CRS is dose-limiting and often necessitates additional interventions, with the IL-6 blocking antibody tocilizumab currently the front-line treatment (Maude et al, 2014) . Pre-clinical studies have also shown that disarming GM-CSF alone using CRISPR inactivation in CAR-T cells or administering blocking antibodies significantly reduces CRS-like symptoms in vitro and in patient-derived xenograft models (Sachdeva et al, 2019;Sterner et al, 2019) . Importantly, two recent phase-I clinical trials (Brudno et al, 2020;Ying et al, 2019) have shown that broadly attenuated CAR-T cell cytokine release can lead to reduced incidence of high-grade CRS and neurotoxicity without compromising the anti-tumour potency of CD19-directed CAR-T cell therapy in B-cell lymphoma patients.…”

The Society for Immunotherapy of Cancer perspective on regulation of interleukin-6 signaling in COVID-19-related systemic inflammatory response

“…http://jitc.bmj.com/ Open access cells dramatically lowers IL-6 and IL-8 secretion 102 and therapeutic inhibition of GM-CSF reduces CAR T-cellassociated CRS in mouse models. 103 Initial evidence from a study of 52 patients with COVID-19 reported from Wuhan, China, elevated levels of circulating GM-CSF were associated with worse clinical outcomes.…”