Abstract 5760: Intra-tissue quantitative analysis using LC/MS will determine the mechanisms of interstitial pneumonia induced by antibody-drug conjugates
Authors: Shigehiro Koganemaru, Hirobumi Fuchigami, Chihiro Morizono, Hiroko Shinohara, Yasutoshi Kuboki, Keiji Furuuchi, Toshimitsu Uenaka, Toshihiko Doi, Masahiro Yasunaga
Journal: Cancer Research (2024)

Abstract Background: Antibody-drug conjugates (ADCs) are a new class of therapeutics that combine a monoclonal antibody with a cytotoxic agent (known as their payloads) via a synthetic linker. While ADCs are one of game changer and are the fastest growing classes with novel clinical benefits recently, interstitial pneumonia (ILD) is a clinical problem as a fatal adverse event. However, the mechanism of ADC-induced ILD is under investigation. Purpose: We clarify ADC-ILD mechanism by analyzing released payload distribution in the lung using a novel Trophoblast cell surface antigen 2 (TROP2)-targeting ADC. Material and Methods: TROP2-eribulin is a novel TROP2-targeting ADC composed of the TROP2 antibody and cytotoxic payload eribulin conjugated by a cleavable linker. Four human cancer cell lines (A549, HeLa, NCI-H2110 and SKOV3) and one human- leukemia monocytic cell line (THP-1) were used in this study. Cell surface TROP2 antigen expression was evaluated by flow cytometry. We investigated the concentration and distribution of the TROP2-eribulin and the payload released from the TROP2-eribulin in tumor and lung tissue, using TROP2 low and high expression human cancer xenograft model that were treated with various doses of TROP2-eribulin. The concentration of erbulin was analyzed by liquid chromatograph mass spectrometry, and tissue distribution of ADCs was assessed by immunofluorescence (IF) assay. Results: In vitro analysis showed that the concentration of intracellularly released eribulin was significantly higher in NCI-H2110 with high TROP2 expression compared to A549 with low TROP2 expression (P &amp;lt; 0.05). In vivo analysis, the concentration of released eribulin in tumor tissue was approximately 10-fold higher in the NIC-H2110 xenograft model than the A549 xenograft model, while analysis of lung tissue showed that TROP2-eribulin was distributed in lung tissue in a dose-dependent manner of TROP2-eribulin regardless of TROP2 expression, with significantly more eribulin released in the high-dose group of TROP2-eribulin than in the other dose groups (P &amp;lt; 0.05). IF assay showed that TROP2-eribuilin localized to alveolar macrophages. In the analysis using THP-1 cell, which mimic macrophage function, the concentration of eribulin released from TROP2-eribuilin was significantly reduced by the use of an Fc receptor inhibitor (P &amp;lt; 0.05). Conclusions: Nonspecific uptake of ADC by alveolar macrophages via Fcγ receptors releases cytotoxic payload into lung tissue, regardless of tumor antigen expression levels. This understanding can help elucidate the pathogenesis of ADC-induced ILD. Citation Format: Shigehiro Koganemaru, Hirobumi Fuchigami, Chihiro Morizono, Hiroko Shinohara, Yasutoshi Kuboki, Keiji Furuuchi, Toshimitsu Uenaka, Toshihiko Doi, Masahiro Yasunaga. Intra-tissue quantitative analysis using LC/MS will determine the mechanisms of interstitial pneumonia induced by antibody-drug conjugates [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5760.