ABL001 (asciminib) Efficiently Targets Transplantable BCR::ABL1 Lymphoid Blast Crisis in the Scl-Tta-p210-BCR::ABL1 Mouse Model

Objectives: This study aimed at investigating the therapeutic efficacy of ABL001 in the ScltTA-p210-BCR::ABL1 transgenic mouse model. ABL001 (asciminib) is a novel allosteric inhibitor of ABL kinase demonstrating its particular efficacy against BCR::ABL1 mutations that emerge during treatment with traditional TKIs. However, the activity and tolerability of ABL001 in a primary mouse model of BCR::ABL1 disease remain unknown. In addition, resistance mutations against ABL001 were observed in the myristoyl pocket of BCR::ABL1, but their emergence has not been analyzed in BCR::ABL1+ murine hematopoietic cells. Importantly, leukemic stem cells (LSCs) expressing BCR::ABL1 have developed mechanisms to evade eradication by tyrosine kinase inhibitors, underscoring the necessity for alternative therapeutic approaches.

Methods: 5-fluorouracil- (5-FU-) treated bone marrow (BM) cells harvested from FVB/N-tg(ScltTA)(tetO- p210-BCR::ABL1) double-transgenic (dtg) donor mice (CD45.1+) were transplanted into lethally irradiated recipient mice (CD45.2+). One week after transplantation, tetracycline (tet) was removed from the drinking water to induce Bcr-Abl1 expression. Following an additional week, mice were treated daily with ABL001 or vehicle control by oral gavage (30 mg/kg body weight). Therapeutic effects were assessed using flow cytometry (peripheral blood, spleen, and BM), histological examination, RNA analysis and DNA sequencing of the myristoyl pocket of BCR::ABL1. Secondary transplants using splenocytes were performed to analyze whether ABL001 treatment was able to suppress BCR::ABL1-positive repopulating malignant stem cells. Additionally, we assessed potential ABL001-resistant subclones after secondary transplantation by subjecting the mice to further ABL001 treatment, modeling clinically relevant mutations in BCR::ABL1-positive disease.

Results: Within 3 weeks after tet withdrawal, the transplanted mice developed an aggressive B-lymphoblastic p210-BCR::ABL1-positive disease, mimicking lymphoid blast crisis, as described previously (Koschmieder et al., Blood, 2005). Interestingly, ABL001 treatment significantly reduced this BCR::ABL1-driven phenotype as indicated by suppression of malignant B cells and restoration of normal hematopoiesis. ABL001 also significantly improved survival compared to the vehicle control group (p=0.026). Histological analysis confirmed the alleviation of the BCR::ABL1-driven phenotype in spleen and BM. Furthermore, ABL001 treatment significantly reduced BCR::ABL1 transcripts to background levels, demonstrating its ability to suppress BCR::ABL1-induced disease in the BM. The efficacy of ABL001 in reducing the malignant clone correlated with normalization of spleen weight. Essentially, ABL001 treatment normalized the phenotypic long-term repopulating hematopoietic stem cell (LT-HSC) population in the BM, suggesting restoration of the stem cell pool and targeting malignant LT-HSCs. This was supported by secondary transplantation experiments, showing that several mice receiving ABL001-treated splenocytes failed to develop leukemia, while all mice who received vehicle-treated cells did. Moreover, none of the secondary transplanted mice who were treated with ABL001 developed leukemia. Interestingly, Sanger sequencing of the BCR::ABL1 myristoyl pocket region demonstrated wildtype sequences and the absence of resistance mutations.

Conclusion: Our study demonstrates the efficacy of ABL001 in ameliorating p210-BCR::ABL1-induced lymphoid blast crisis by specifically targeting BCR::ABL1-positive malignant stem cells and restoring normal hematopoiesis in a transgenic mouse model. These promising findings highlight the potential of ABL001 as an alternative therapeutic strategy for first-line treatment of BCR::ABL1-driven leukemia, including B-lymphoblastic blast crisis. Furthermore, since TKI treatment alone has not proven successful in patients with lymphoid blast crisis, this now opens new avenues for the in vivo-testing of novel combination treatments of ABL001 with chemotherapy or other targeting agents.