Berger Marc G, Lebecque Benjamin, Tassin Thomas, Dannus Louis-Thomas, Berger Juliette, Soucal Mélanie, Guerci Agnès, Cony-Makhoul Pascale, Johnson Hyacinthe, Etienne Gabriel, Guyotat Denis, Gagnieu Marie-Claude, Pereira Bruno, Saugues Sandrine, Tournilhac Olivier, Hermet Eric, Bourgne Céline
Hématologie Biologique, CHU Clermont-Ferrand, Hôpital Estaing, 1 place Lucie et Raymond Aubrac, 63003, Clermont-Ferrand Cedex 1, France.
Equipe d'Accueil 7453 CHELTER, Université Clermont Auvergne, CHU Clermont-Ferrand, Hôpital Estaing, 1 place Lucie et Raymond Aubrac, 63003, Clermont-Ferrand Cedex 1, France.
Sci Rep. 2021 Mar 17;11(1):6187. doi: 10.1038/s41598-021-85734-0.
Accumulation in target cells is an essential pharmacokinetic step of targeted therapies. Tyrosine Kinase Inhibitors (TKI) against the BCR-ABL fusion protein in Chronic Phase-Chronic Myeloid Leukaemia (CP-CML) cells constitute a unique model in terms of efficacy, specificity, and in vivo demonstration of response heterogeneity by target cells. The overall therapeutic response to nilotinib is heterogeneous with no satisfactory explanation. To better understand the patients' response heterogeneity, we quantified nilotinib uptake by primary CP-CML cells in standardized conditions using flow cytometry, which allowed also distinguishing mature (polymorphonuclear cells) from immature (CD34) cells. Nilotinib was undetectable in 13.3% of PMN and 40% of CD34 cells. Moreover, in CD34 cells, intracellular nilotinib did not completely abolish BCR-ABL activity (monitored by CrkL phosphorylation inhibition), although nilotinib accumulated well in most CD34 cell samples. Intracellular nilotinib concentration was inversely correlated with disease burden parameters, Sokal score, and early haematologic response at day 6 ± 1 only in PMN, suggesting an intrinsic ability to limit nilotinib entry in the forms with higher tumor cell burdenat diagnosis. These findings suggest that nilotinib accumulation in CP-CML cells is influenced by individual characteristics and intra-clonal heterogeneity, and might be used for pharmacokinetic studies and to assess the therapeutic response.
在靶细胞中蓄积是靶向治疗重要的药代动力学步骤。针对慢性期慢性髓性白血病（CP-CML）细胞中BCR-ABL融合蛋白的酪氨酸激酶抑制剂（TKI）在疗效、特异性以及靶细胞体内反应异质性的展示方面构成了一个独特的模型。对尼罗替尼的总体治疗反应存在异质性，且尚无令人满意的解释。为了更好地理解患者的反应异质性，我们在标准化条件下使用流式细胞术对原代CP-CML细胞摄取尼罗替尼的情况进行了定量，这也使得能够区分成熟（多形核细胞）和未成熟（CD34）细胞。在13.3%的多形核细胞和40%的CD34细胞中未检测到尼罗替尼。此外，在CD34细胞中，尽管尼罗替尼在大多数CD34细胞样本中蓄积良好，但细胞内尼罗替尼并未完全消除BCR-ABL活性（通过CrkL磷酸化抑制进行监测）。仅在多形核细胞中，细胞内尼罗替尼浓度与疾病负担参数、索卡尔评分以及第6±1天的早期血液学反应呈负相关，这表明在诊断时肿瘤细胞负担较高的形式中存在限制尼罗替尼进入的内在能力。这些发现表明，CP-CML细胞中尼罗替尼的蓄积受个体特征和克隆内异质性的影响，可能用于药代动力学研究和评估治疗反应。
