Relocalization of KIT D816V to Cell Surface After Dasatinib Treatment: Potential Clinical Implications

Systemic mastocytosis is a disease associated with a D816V mutation on KIT tyrosine kinase receptor KIT in mastocytes. We describe the treatment of blood samples from 28 patients with systemic mastocytosis with the tyrosine kinase inhibitor dasatinib, which enhances KIT D816V flow cytometry detection on mast cell-related cells, with a more pronounced increase on the severe forms of the disease. Background: Systemic mastocytosis (SM) is a heterogeneous disease that displays variable aggressivity. Adults with SM frequently have a D816V mutation in the tyrosine kinase (TK) receptor gene KIT. We previously reported that, in a Chinese hamster ovarian cell model expressing exogenous KIT variants, constitutive activating KIT mutations induced intracellular mislocalization of KIT reversed by inhibition of KIT TK activity. Hence, we hypothesized that inhibition of KIT kinase activity by the TK inhibitor dasatinib could be useful to increase KIT detection sensitivity in samples from patients with SM. Patients And Methods: We tested this hypothesis on a BaF/3 cell line modified to express either KIT wild-type (WT) or KIT D816V, on the human mastocytoma cell line HMC1.2, and among 28 patients with proven SM who did (n = 24) or did not (n = 4) carry the D816V KIT mutation and displayed various SM subtypes by using a simple flow cytometry assay to quantify KIT relocalization upon dasatinib treatment. Results: We confirm KIT cell surface increase upon dasatinib treatment on BaF/3 KIT D816V and HMC1.2 cell lines but not on BaF/3 KIT WT cell line. The analysis of bone marrow and peripheral blood samples of patients with SM showed KIT surface level increase for patients with the KIT D816V mutation but not for patients who had no KIT mutation. Interestingly, the extent of KIT level relocalization correlates with SM severity, with a higher relocalization for patients with aggressive forms compared with indolent forms. Conclusions: Overall, results of this study suggests that treating the peripheral blood sample with dasatinib of a patient with SM before analysis by flow cytometry could contribute to narrowing the SM diagnosis. Clinical Lymphoma, Myeloma & Leukemia, Vol. 13, No. 1, 62-9 (c) 2013 Elsevier Inc. All rights reserved.