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

  • J Enzyme Inhib Med Chem. 2026 Dec;41(1):2638027. doi: 10.1080/14756366.2026.2638027.
Wariya Nirachonkul  1 ,  Mark P Farrell  2 ,  Thomas J Tolbert  3 ,  Siriporn Okonogi  4  5 ,  Singkome Tima  1  5  6 ,  Songyot Anuchapreeda  1  5  6 ,  Sawitree Chiampanichayakul  1  5  6 ,  Teruna J Siahaan  3
Affiliations
  • 1. Department of Medical Technology, Chiang Mai University, Chiang Mai, Thailand.
  • 2. Department of Medicinal Chemistry, The University of Kansas, Lawrence, KS, USA.
  • 3. Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence, KS, USA.
  • 4. Department of Pharmaceutical Sciences, Chiang Mai University, Chiang Mai, Thailand.
  • 5. Center of Excellence in Pharmaceutical Nanotechnology, Chiang Mai University, Chiang Mai, Thailand.
  • 6. Cancer Research Unit of Associated Medical Sciences (AMS CRU), Chiang Mai University, Thailand.
PMID: 41784256 DOI: 10.1080/14756366.2026.2638027
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.

Keywords
FLT3-MMAE conjugate; FLT3-internal tandem duplication (FLT3-ITD); MV4-11; acute myeloblastic leukaemia (AML); antibody-drug conjugates (ADCs).
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