Endothelium as a mediator of BCR-ABL tyrosine kinase inhibitor vascular toxicity

靶向bcr-abl的酪氨酸激酶抑制剂(TKIs)转化慢性粒细胞白血病(CML) 从预后不良的致命疾病到预期寿命接近的可控慢性病 导致慢性粒细胞白血病患病率增加，预计将继续 至少要上升到2050年。这一成功是由具有更高效力的新一代BCR-ABL TKI推动的。 然而，一些BCR-ABL TKI，最著名的是达沙替尼、波拉替尼和尼洛替尼，已经与 心血管毒性包括增加动脉血栓形成事件的风险，如冠状动脉血栓形成， 外周动脉闭塞和脑血管意外。我们的实验室之前帮助开发了一种蛋白质组 鉴定人脐静脉内皮细胞(HUVECs)磷酸化蛋白质组特征的方法 BCR-ABL TKIs治疗可预测临床血管毒性并与内皮细胞(EC)相关 体外功能障碍。ECS排列血管管腔，保护血管系统免受病理性血小板粘连和 血栓形成。相反，损害内皮细胞愈合能力的内皮细胞功能障碍也暴露于血栓形成因子。 在基底膜上，随着EC促凝血蛋白的上调，还没有被探索 BCR-ABL TKI毒性可能导致动脉血栓形成风险增加的机制。阿西米尼， 一种新的变构BCR-ABL抑制剂扩大了CML的治疗范围，并于2021年获得 FDA批准之前两次BCR-ABL TKI失败的患者。阿司米尼的心血管安全性研究进展 而且它对体外培养的内皮细胞的影响尚不清楚，对bcr-abl TKI血管也没有缓解策略。 毒性。来自我们实验室的新的初步数据支持，阿斯米尼的毒性低于波纳替尼、达沙替尼和 尼洛替尼在HUVEC划痕中的体外愈合试验表明，尼洛替尼损伤小鼠颈动脉内皮细胞 细胞创伤在体内愈合，以及磷酸化蛋白质组筛选可以识别出“逆转”有毒的bcr的药物。 BCR-ABL TKI暴露后的ABL-TKI磷酸化蛋白质组学特征和提高EC存活率。我们建议 用体外试验比较阿司米尼与伊马替尼(无毒BCR-ABL TKI)、波拉替尼、阿司匹林的EC毒性。 尼洛替尼和达沙替尼(均已知对内皮细胞有毒性)，并探讨这些化合物在体内的EC毒性。 一种小鼠颈动脉钢丝损伤模型。我们还建议使用蛋白质组学来确定磷酸化- 经阿司米尼处理的人脐静脉内皮细胞蛋白质组图谱，筛选批准的心脏代谢药物的能力 “逆转”有毒的bcr-abl TKIs的磷酸化蛋白质组EC特征，并测试目标药物是否 经我们的筛选证实，在体外和体内都能逆转EC的毒性。这项建议中的研究结果将 提供有关治疗慢性粒细胞白血病的新疗法阿西米尼的潜在血管毒性的有价值的信息，提供 更全面地了解当前BCR-ABL TKIs的血管毒性，并迅速识别 治疗或预防慢性粒细胞白血病幸存者血管毒性的可翻译药物。

Tyrosine kinase inhibitors (TKIs) that target BCR-ABL have transformed Chronic Myelogenous Leukemia (CML) from a deadly condition with a poor prognosis to a manageable chronic disease with a life expectancy close to that of the normal population, resulting in an increase in the prevalence of CML that is expected to continue to rise until at least 2050. This success is driven by newer generations of BCR-ABL TKIs with improved potency. Some BCR-ABL TKIs however, most notably dasatinib, ponatinib and nilotinib, have been associated with cardiovascular toxicity including increased risk of arterial thrombotic events such as coronary artery thrombosis, peripheral arterial occlusion, and cerebrovascular accidents. Our lab previously helped develop a proteomic approach which identified a phospho-proteomic signature of human umbilical vein endothelial cells (HUVECs) treated with BCR-ABL TKIs that predicts clinical vascular toxicity and correlates with endothelial cell (EC) dysfunction in vitro. ECs line the vessel lumen and protect the vasculature from pathologic platelet adhesion and thrombosis. Conversely, EC dysfunction that impairs the ability of ECs to heal also exposes prothrombotic factors on the basement membrane that, along with EC upregulation of pro-coagulant proteins, are unexplored mechanisms by which BCR-ABL TKI toxicity could contribute to increased risk of arterial thrombosis. Asciminib, a novel allosteric BCR-ABL inhibitor, has expanded the landscape of treatment in CML and in 2021 received FDA approval for patients who have failed two prior BCR-ABL TKIs. The cardiovascular safety profile of asciminib and its effects on ECs in vitro is unknown, nor are mitigating strategies available for BCR-ABL TKI vascular toxicity. New preliminary data from our lab support that asciminib is less toxic than ponatinib, dasatinib, and nilotinib in a HUVEC scratch would healing assay in vitro, that ponatinib impairs mouse carotid artery endothelial cell wound healing in vivo, and that phospho-proteomic screening can identify drugs that “reverse” the toxic BCR- ABL TKI phospho-proteomic signature and improve EC survival after BCR-ABL TKI exposure. We propose to use in vitro assays to compare the EC toxicity of asciminib to imatinib (non-toxic BCR-ABL TKI), ponatinib, nilotinib, and dasatinib (all known to be toxic to ECs), and explore the EC toxicity of these compounds in vivo in a mouse carotid artery wire injury model. We also propose to use proteomics to determine the phospho- proteomic profile of HUVECs treated with asciminib, screen approved cardiometabolic drugs for their ability to “reverse” the phospho-proteomic EC signature of toxic BCR-ABL TKIs, and test whether the target drugs identified by our screen can reverse EC toxicity in vitro and in vivo. The results of the studies in this proposal will provide valuable information about the potential vascular toxicity of asciminib, a novel therapy for CML, provide a more comprehensive understanding of the vascular toxicity of current BCR-ABL TKIs, and identify rapidly translatable agents to treat or prevent vascular toxicity in CML survivors.