Tumour-associated macrophages process drug and radio-conjugates of the dead tumour cell-targeting APOMAB® antibody

APOMAB (chDAB4) is a dead tumour cell-targeting antibody which has been used preclinically as a diagnostic agent and therapeutically as a radioimmunotherapy and antibody drug conjugate (ADC). However, little is known of the intra-tumour processing of chDAB4 when bound to dead tumour cells. In this study we examine the role of macrophages in the in vitro and in vivo processing of radiolabelled chDAB4 and a chDAB4 ADC. We found that chDAB4 binds to macrophages in vitro, resulting in the killing of macrophages when using the ADC, chDAB4-SG3249. Free drug released by the macrophage processing of chDAB4-SG3249 could result in killing of 'bystander' Lewis lung (LL2) carcinoma cells. Furthermore, macrophages phagocytosed chDAB4-bound dead LL2 cells and were killed when they phagocytosed chDAB4-SG3249-bound dead LL2 cells in vitro. In vivo, we found markedly different tumour retention of chDAB4 in the LL2 tumour model depending on whether it was radiolabelled with a residualising radionuclide (⁸⁹Zr), which is retained intracellularly, or a non-residualising radionuclide (¹²⁴I), which can diffuse out of the cell. This prolonged retention of ⁸⁹Zr vs¹²⁴I indicated intra-tumoral processing of chDAB4 in vivo. The tumour uptake of ⁸⁹Zr-chDAB4 was reduced after macrophage depletion, which also reduced the efficacy of the chDAB4 ADC in vivo. This study shows that macrophages can process chDAB4 and chDAB4 ADC in vitro and shows the importance of tumour-associated macrophages in the tumour retention of chDAB4 and the efficacy of chDAB4 ADC in vivo.

Keywords: APOMAB; Antibody drug conjugate; Bystander killing; Iodine-124; La/SSB; Macrophages; PBD dimer; Zirconium-89.