A patient-derived lung-on-chip model to evaluate on-target/off-tumour toxicity of the therapeutic FOLR1-targeting T-Cell Bispecific antibody

Abstract

The overexpression of Folate-receptor 1 (FOLR1) in various types of cancer (ovarian, lung, renal), makes it an attractive candidate for targeted-tumour immunotherapy. T-cell bispecific antibodies are engineered to recognize both FOLR1 and the T-cell receptor CD3 (FOLR1-TCB), enhancing tumour recognition, assault and killing by T-cells. Despite their therapeutic potential, such antibodies present the risk of on-target/off-tumour toxicity, as FOLR1 is also expressed in healthy epithelial cells in the lung or the kidney.

In this work, FOLR1-TCB pulmonary toxicity was evaluated using an advanced lung-on-chip in-vitro model. With that purpose, human epithelial and/or endothelial barrier models were treated with FOLR1-TCB in the presence of peripheral blood mononuclear cells (PBMC). According to our results, FOLR1-TCB induced a pronounced on-target/off-tumour damage in our alveolar model as indicated by increased cytotoxicity, barrier leakage (TER) and pro-inflammatory cytokine release (e.g., IL-6, Granzyme B). Additionally, FOLR1-TCB treatment induced T-cell specific activation, as observed by flow cytometry analysis, as well as targeted attachment to the epithelium, as detected by live imaging tracking.

Taken together, our results suggest that our patient-derived alveolar lung-on-chip model can successfully predict off-tumour TCB adverse effects. This highlights the importance of considering the immune component, allowing cell-cell interactions, and providing an in-vivo-like microenvironment for relevant safety evaluation of therapeutic antibodies during the preclinical phase of drug development.