Dissecting the Mechanisms Underlying the Cytokine Release Syndrome (CRS) Mediated by T-Cell Bispecific Antibodies

Abstract

Purpose: Target-dependent TCB activity can result in strong and systemic release of cytokines that may develop into Cytokine Release Syndrome (CRS), highlighting the need to understand and prevent this complex clinical syndrome. Experimental design: We explored the cellular and molecular players involved in TCB-mediated cytokine release by single cell RNA sequencing of whole blood treated with CD20-TCB together with bulk RNA sequencing of endothelial cells exposed to TCB-induced cytokine release. We used the in vitro whole blood assay and an in vivo DLBCL model in immunocompetent humanized mice to assess the effects of dexamethasone, anti-TNF-a, anti-IL-6R, anti-IL-1R and inflammasome inhibition, on TCB-mediated cytokine release and anti-tumor activity. Results: Activated T cells release TNF-α, IFN-γ, IL-2, IL-8 and MIP-1β, which rapidly activate monocytes, neutrophils, DCs and NKs along with surrounding T cells to amplify the cascade further, leading to TNF-α, IL-8, IL-6, IL-1β, MCP-1, MIP-1α, MIP-1β and IP-10 release. Endothelial cells contribute to IL-6 and IL-1β release and at the same time release several chemokines (MCP-1, IP-10, MIP-1α and MIP-1β). Dexamethasone and TNF-α blockade efficiently reduced CD20-TCB-mediated cytokine release while IL-6R blockade, inflammasome inhibition and IL1-R blockade induced a less pronounced effect. Dexamethasone, IL-6R blockade, IL-1R blockade and the inflammasome inhibitor did not interfere with CD20-TCB activity, in contrast to TNF-α blockade, which partially inhibited anti-tumor activity. Conclusion: Our work sheds a new light on the cellular and molecular players involved in cytokine release driven by TCBs and provides rationale for the prevention of CRS in patients treated with TCBs.

Development of pharmacological immunoregulatory anti-cancer therapeutics: current mechanistic studies and clinical opportunities

“…It is a systemic inflammatory response with symptoms ranging from fever, fatigue, and headache to multiorgan failure, triggered by T cell activation, with myeloid cells and TNF-α being the main mediators of the systemic cytokine release. 356 , 357 To advance the further application of TCEs, the management of their using and the handling of adverse events should be improved, for example, with stepwise dosing, properly using tocilizumab, corticosteroids, or TNF-α blockade, and supportive 353 – 355 , 358 care. Regarding the induction of ADAs, increased engineering and artificial design may result in greater differences between bsAbs and endogenous immunoglobulins, and bsAbs could therefore potentially contain new epitopes that elevate antigenicity and subsequently increase the likelihood of ADA development.…”

The enhanced antitumor activity of bispecific antibody targeting PD-1/PD-L1 signaling

“…Predominantly, these challenges include managing adverse reactions associated with treatment, mitigating both on-target and off-target toxicities, and navigating the intricacies of the immunosuppressive TME [ 41 ]. A critical issue associated with BsAbs, especially those with intact Fc domains, is the risk of off-target toxicity, exemplified by Cytokine Release Syndrome (CRS) [ 45 ]. CRS is a systemic inflammatory reaction characterized by a spectrum of clinical manifestations, from mild symptoms to severe, potentially fatal conditions, often marked by laboratory signs such as cytopenia [ 46 ].…”

Immune‐related adverse events of antibody‐based biological medicines in cancer therapy

“…Recent research sheds light on critical aspects of CRS and provides insights for clinical management of CRS following BsAb treatment. 108 Using single‐cell RNA and bulk RNA sequencing, the researchers identified T‐cells as initial drivers of cytokine and chemokine release, activating a myeloid‐cell‐coordinated downstream cascade of events. These studies identify the significant role of endothelial cells and neutrophil‐derived mediators as key contributors to inflammatory response elicited by BsAb such as blinatumomab.…”