Toledo Marcelo A S, Gatz Malrun, Sontag Stephanie, Gleixner Karoline V, Eisenwort Gregor, Feldberg Kristina, Hamouda Ahmed E I, Kluge Frederick, Guareschi Riccardo, Rossetti Giulia, Sechi Antonio S, Dufva Olli M J, Mustjoki Satu M, Maurer Angela, Schüler Herdit M, Goetzke Roman, Braunschweig Till, Kaiser Anne, Panse Jens, Jawhar Mohamad, Reiter Andreas, Hilberg Frank, Ettmayer Peter, Wagner Wolfgang, Koschmieder Steffen, Brümmendorf Tim H, Valent Peter, Chatain Nicolas, Zenke Martin
Institute for Biomedical Engineering, Department of Cell Biology, RWTH Aachen University Medical School, Aachen, Germany.
Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University, Aachen, Germany.
Blood. 2021 Apr 15;137(15):2070-2084. doi: 10.1182/blood.2019004509.
The KIT D816V mutation is found in >80% of patients with systemic mastocytosis (SM) and is key to neoplastic mast cell (MC) expansion and accumulation in affected organs. Therefore, KIT D816V represents a prime therapeutic target for SM. Here, we generated a panel of patient-specific KIT D816V induced pluripotent stem cells (iPSCs) from patients with aggressive SM and mast cell leukemia to develop a patient-specific SM disease model for mechanistic and drug-discovery studies. KIT D816V iPSCs differentiated into neoplastic hematopoietic progenitor cells and MCs with patient-specific phenotypic features, thereby reflecting the heterogeneity of the disease. CRISPR/Cas9n-engineered KIT D816V human embryonic stem cells (ESCs), when differentiated into hematopoietic cells, recapitulated the phenotype observed for KIT D816V iPSC hematopoiesis. KIT D816V causes constitutive activation of the KIT tyrosine kinase receptor, and we exploited our iPSCs and ESCs to investigate new tyrosine kinase inhibitors targeting KIT D816V. Our study identified nintedanib, a US Food and Drug Administration-approved angiokinase inhibitor that targets vascular endothelial growth factor receptor, platelet-derived growth factor receptor, and fibroblast growth factor receptor, as a novel KIT D816V inhibitor. Nintedanib selectively reduced the viability of iPSC-derived KIT D816V hematopoietic progenitor cells and MCs in the nanomolar range. Nintedanib was also active on primary samples of KIT D816V SM patients. Molecular docking studies show that nintedanib binds to the adenosine triphosphate binding pocket of inactive KIT D816V. Our results suggest nintedanib as a new drug candidate for KIT D816V-targeted therapy of advanced SM.
在超过80%的系统性肥大细胞增多症（SM）患者中发现了KIT D816V突变，该突变是肿瘤性肥大细胞（MC）在受累器官中扩增和聚集的关键。因此，KIT D816V是SM的主要治疗靶点。在此，我们从侵袭性SM和肥大细胞白血病患者中生成了一组患者特异性的KIT D816V诱导多能干细胞（iPSC），以建立用于机制和药物发现研究的患者特异性SM疾病模型。KIT D816V iPSC分化为具有患者特异性表型特征的肿瘤性造血祖细胞和MC，从而反映了疾病的异质性。经CRISPR/Cas9n工程改造的KIT D816V人类胚胎干细胞（ESC）在分化为造血细胞时，重现了KIT D816V iPSC造血所观察到的表型。KIT D816V导致KIT酪氨酸激酶受体的组成性激活，我们利用我们的iPSC和ESC来研究靶向KIT D816V的新型酪氨酸激酶抑制剂。我们的研究确定了尼达尼布，一种美国食品药品监督管理局批准的血管激酶抑制剂，其靶向血管内皮生长因子受体、血小板衍生生长因子受体和成纤维细胞生长因子受体，作为一种新型的KIT D816V抑制剂。尼达尼布在纳摩尔范围内选择性降低了iPSC衍生的KIT D816V造血祖细胞和MC的活力。尼达尼布对KIT D816V SM患者的原代样本也有活性。分子对接研究表明，尼达尼布与无活性KIT D816V的三磷酸腺苷结合口袋结合。我们的结果表明，尼达尼布是晚期SM的KIT D816V靶向治疗的新候选药物。
