Modeling Tyrosine Kinase Inhibitor-Induced Vascular Dysfunction Using Human iPSCs

项目摘要 酪氨酸激酶抑制剂（TKI）已被证明可以显着减少各种恶性相关的肿瘤， 死亡率在过去的二十年里。然而，由于其潜在的血管毒性， 可能导致高血压、心肌梗塞、中风和外周动脉疾病。尽管有这些 安全性问题，TKI诱导的血管毒性（TKI-VT）的潜在机制知之甚少。到 为了克服这一挑战，我们建议利用人类iPSC，最先进的多组学方法， CRISPR筛选以研究TKI-VT的分子和细胞机制并确定可药物靶点 可以在动物模型中进一步测试。具体而言，在目标1中，我们将全面描述人类诱导的 多能干细胞衍生的心脏周细胞（iPSC-PC）是一种重要但很少探索的心脏细胞类型， 定义TKI-VT的细胞机制。在目标2中，我们将评估TKI如何诱导中断的细胞串扰 iPSC-PC和iPSC衍生的内皮细胞（iPSC-EC）之间的关系， 片上容器（VoC）模型。最后，在目标3中，我们将对TKI处理的iPSC-PC进行CRISPR筛选， iPSC-EC以鉴定潜在的可药物靶点并验证其在小鼠中的治疗功效。成功 这些研究的完成将导致对TKI-VT发病机制的新的机制见解，并有助于开发 有希望的治疗策略，可以预防和/或治疗癌症患者的TKI-VT。此外，该提案 将有助于确定TKI在血管病理生理学中的作用，这可能具有广泛的科学和临床意义。 影响超出了心脏肿瘤学。

PROJECT SUMMARY Tyrosine kinase inhibitors (TKIs) have been shown to significantly decrease a variety of malignancy-related mortality in the past two decades. However, concerns have been raised due to their potential vascular toxicity that could lead to hypertension, myocardial infarction, stroke, and peripheral arterial diseases. Despite these safety concerns, the mechanisms underlying TKI-induced vascular toxicity (TKI-VT) are poorly understood. To overcome this challenge, we propose to leverage human iPSCs, state-of-the-art multi-omics methods, and CRISPR screening to investigate molecular and cellular mechanisms of TKI-VT and identify druggable targets that can be further tested in animal models. Specifically, in Aim 1, we will comprehensively profile human-induced pluripotent stem cell-derived cardiac pericytes (iPSC-PCs), an important but rarely explored cardiac cell type, to define cellular mechanisms of TKI-VT. In Aim 2, we will evaluate how TKIs induce disrupted cellular crosstalk between iPSC-PCs and iPSC-derived endothelial cells (iPSC-ECs) by performing integrative omics on a 3D vessel-on-chip (VoC) model. Finally, in Aim 3, we will perform CRISPR screening on TKI-treated iPSC-PCs and iPSC-ECs to identify potential druggable targets and validate their therapeutic efficacy in mice. Successful completion of these studies will lead to novel mechanistic insights into TKI-VT pathogenesis and help develop promising therapeutic strategies that can prevent and/or treat TKI-VT in cancer patients. Moreover, this proposal will help define the role of TKIs in vascular pathophysiology, which may have broad scientific and clinical implications beyond cardio-oncology.