Novel strategies to understand, predict, and prevent vascular toxicity of targeted CML therapies

分子靶向的癌症治疗，如Abl靶向酪氨酸激酶抑制剂(TKIs)，有 将包括慢性粒细胞白血病(CML)在内的许多晚期癌症转化为慢性病。 尽管采取了有针对性的方法，但血管毒性已经出现。具体地说，第一代CML TKI伊马替尼是安全的，但新一代TKI经常用于耐药CML(尼洛替尼，达沙替尼， 帕纳替尼)使动脉血栓形成的风险增加2-5倍，导致心肌梗死、中风或肢体缺血。 在药物批准之前，没有经过验证的模型来测试血管毒性，因此确定了这种副作用。 只有当不良事件发生并一旦被认识到时，才没有数据指导临床医生如何 以防止这种情况或治疗这些癌症幸存者。这项建议通过以下方式解决了这些知识差距 利用CML TKIs，这是一种既有安全成员又有有毒成员的药物类别。我们和其他人证明了 在一些体外模型中，有毒的CML TKI损伤内皮细胞，并增强小鼠的动脉粥样硬化。作为CML 有潜在心血管危险因素的患者容易发生毒性，我们假设有毒的慢性粒细胞白血病 药物损害特定的内皮细胞(EC)功能，导致斑块发展，炎症， 破裂、血栓形成和缺血。由于这些药物是激酶抑制剂，我们进一步假设 是由信号蛋白的磷酸化状态以一种有害的方式调节引起的 用蛋白质组学的方法证明了这一点。因此，我们现在建议测试 假设有毒的CML TKI作用于内皮细胞，破坏屏障完整性，促进白细胞转运，速度缓慢 伤口愈合，增加与血小板的相互作用，从而促进动脉粥样硬化表型 容易破裂和血栓形成，药物诱导的EC蛋白质组图谱可以预测血管 毒性，并确定减轻处理方法。我们通过使用多种创新方法的两个目标对此进行测试： SA1使用人类工程微血管(HEMV)询问每个CML TKI的影响，以 在小鼠体内检测对血管通透性和血小板聚集性的影响 显微镜观察白细胞与内皮细胞的相互作用，颈动脉线损伤测量血管再狭窄 内皮化，以及使用FACS的Apo-E-KO小鼠来量化动脉粥样硬化中的血管炎症。 SA2使用基于靶向质谱学的磷酸蛋白质组分析来描述新出现的 CML TKI和一系列被批准用于治疗人内皮细胞心血管疾病的药物以确定这是否可以 预测新的TKIs的毒性并确定缓解疗法。预测的毒性和“解毒剂” 反对有毒的蛋白质组特征将使用SA1中描述的体外和体内模型进行测试。 AIMS的完成将通过验证预测血管的临床前模型来改变心脏肿瘤学 慢性粒细胞白血病TKIs的安全性，并确定可迅速转化为 心脏肿瘤护理。

Molecularly targeted cancer therapies, such as Abl-targeted tyrosine kinase inhibitors (TKIs), have converting many terminal cancers, including chronic myeloid leukemia (CML), into chronic diseases. Despite the targeted approach, vascular toxicities have emerged. Specifically, the first-generation CML TKI imatinib is safe, yet newer generation TKIs frequently used for resistant CML (nilotinib, dasatinib, ponatinib) confer a 2-5 fold increased risk of arterial thrombosis causing MI, stroke, or limb ischemia. With no validated models to test for vascular toxicity prior to drug approval, this side effect was identified only when adverse events accrued and once recognized, there are no data to guide clinicians as to how to prevent this or to treat these cancer survivors. This proposal addresses these gaps in knowledge by leveraging the CML TKIs, a drug class with both safe and toxic members. We and others showed that toxic CML TKIs damage ECs in some in vitro models and enhance atherosclerosis in mice. As CML patients with underlying CV risk factors are predisposed to toxicity, we hypothesize that the toxic CML drugs impair specific endothelial cell (EC) functions that lead to plaque development, inflammation, rupture, thrombosis and ischemia. As these drugs are kinase inhibitors, we further posit that the toxicity is caused by modulation of the phosphorylation state of signaling proteins in a manner that is deleterious to EC function and have shown this using proteomic approaches. Thus, we now propose to test the hypothesis that toxic CML TKIs act on ECs to impair barrier integrity, enhance leukocyte trafficking, slow wound healing, and increase interaction with platelets, thereby promoting an atherosclerosis phenotype prone to rupture and thrombosis, and that the drug-induced EC proteomic profile can predict vascular toxicity and identify mitigating treatments. We test this with 2 aims using multiple innovative approaches: SA1 interrogates the impact of each CML TKI using human engineered microvessels (hEMVs) to examine the impact on vascular permeability and platelet aggregation and in mice using intravital microscopy to quantify leukocyte-EC interaction, carotid wire injury to measure vascular re- endothelialization, and Apo-E-KO mice with FACS to quantify vascular inflammation in atherosclerosis. SA2 uses a targeted mass spectrometry based phosphoproteomic assay to profile the effects of emerging CML TKIs and a broad range of drugs approved for CV disease in human ECs to determine if this can predict toxicity of new TKIs and identify mitigating therapies. Predicted toxicities and “antidotes” that oppose the toxic proteomic signature will be tested using the in vitro and in vivo models described in SA1. Completion of the aims will transform cardiooncology by validating preclinical models to predict vascular safety of CML TKIs and identify potential treatments for vascular toxicity that can be rapidly translated to cardiooncology care.