# Mislocalisation of FLT3-ITD receptor contributes to MV4-11 leukaemia cell resistance to antibody-drug conjugate

## TL;DR

The study shows that FLT3-ITD leukaemia cells resist antibody-drug conjugates due to mislocalization of the receptor, which affects drug delivery.

## Contribution

The novel contribution is identifying how FLT3 trafficking differences impact ADC efficacy in leukaemia cells.

## Key findings

- FLT3 mAb traffics to lysosomes in FLT3-wt cells but accumulates in the Golgi in FLT3-ITD cells.
- Anti-FLT3 mAb-MMAE showed lower cytotoxicity in MV4-11 cells due to impaired lysosomal trafficking.
- Effective lysosomal targeting is essential for ADC activity in FLT3-targeted therapies.

## Abstract

FMS-like tyrosine kinase 3 (FLT3/CD135) regulates haematopoiesis and is frequently mutated as FLT3-internal tandem duplication (FLT3-ITD) in acute myeloid leukaemia (AML), associated with poor prognosis. Although FLT3 inhibitors show clinical benefits, resistance remains a challenge. This study hypothesises that antibody-drug conjugate (ADC) efficacy depends on distinct FLT3 trafficking mechanisms in FLT3-wt and FLT3-ITD cells. Confocal imaging showed that in THP-1 (FLT3-wt) cells, FLT3 mAb trafficked to lysosomes, while in MV4-11 (FLT3-ITD) cells, it accumulated in the Golgi. To evaluate the impact of this trafficking difference, we synthesised an anti-FLT3 mAb-MMAE, linked via a Val-Cit-PAB linker at the Fc N-glycan, which exhibited lower cytotoxicity in MV4-11 than THP-1 cells, indicating that the impaired lysosomal trafficking of FLT3-ITD limits drug release and reduces ADC potency. These findings highlight that effective lysosomal targeting is essential for ADC activity and suggest that optimising linker design or restoring lysosome trafficking may enhance FLT3-targeted ADC in AML.