Unraveling the role of endothelium in chemotherapy-induced cardiotoxicity

项目总结/摘要 多柔比星是一种高效的化疗药物，常用于治疗多种癌症，但它的用途是 由于心脏毒性而受到限制。心脏毒性包括无症状的左心室射血减少 严重症状性心力衰竭（III至IV级）。急性阿霉素诱导的心脏毒性（DIC） 约11%的患者发生，约36%的患者可发生长期心脏毒性副作用， 治疗后数年。尽管它是最有效的一类抗癌药物，并且自过去五年以来被广泛使用， 几十年来，DIC的分子机制仍然知之甚少。到目前为止，三大国际... 已经提出了多柔比星的心脏毒性作用的相关机制：（i）产生反应性 氧物种（ROS）和随后的膜损伤，（ii）抑制拓扑异构酶II-β（TOP 2B） 线粒体拓扑异构酶I（TOP1 MT），和（iii）调节细胞内钙释放。但随着 DIC患者的心脏毒性可能在几年或几十年内不会出现，更好地了解不同的 DIC的机制在不同的心脏细胞类型和他们的串扰可以有显着的影响， 寻找治疗方法 内皮细胞是心血管系统的重要组成部分，对CM形成保护屏障 并释放旁分泌因子以维持CM健康和功能。已经表明，DOX破坏了 正常的内皮生理功能，可导致严重的慢性血管 例如动脉粥样硬化等疾病，这通常会导致心脏功能障碍。与知识同行而 功能失调的EC可以对CM功能产生负面影响，我们需要更好地了解整体 内皮细胞在阿霉素诱导的心肌损伤中的作用。尽管取得了令人印象深刻的进展， 人们已经注意到EC和CM之间的细胞间信号传导的潜在重要性， 事实上，EC起到旁分泌功能以增强CM中的信号传导，特别是在药理学背景下， 刺激.这种知识差距阻碍了我们对器官功能障碍的全面理解， 细胞水平。我们建议的总体目标是使用多学科的方法，将人类 iPSC、生物工程工具和NGS，以获得对DIC发病机制的新见解。我们将追捕三名 具体目标。目的一：建立一个研究内皮细胞在DIC中作用的实验平台。为此我们 将重现DIC患者iPSC衍生细胞与3D工程心脏组织的EC-CM串扰 （EHTs）。在目标2中：我们将使用单克隆抗体来破译用DOX处理的EHT中EC-CM串扰的机制。 细胞方法（scRNA-seq和scATAC-seq）。在目标3中：我们将验证EC-CM的关键监管参与者 在DIC动物模型中的串扰。我们的建议得到了来自多个国家的令人信服的初步数据的支持， 调查员纪律小组。我们相信，我们有能力在五年内实现项目目标。 年

Project Summary/Abstract Doxorubicin is a highly effective chemotherapy drug commonly used to treat multiple cancers, but its use is limited due to cardiotoxicity. Cardiotoxicity can range from asymptomatic reduction in left ventricular ejection fraction to highly symptomatic heart failure (Class III to IV). Acute doxorubicin-induced cardiotoxicity (DIC) occurs in ~11% of patients, and long-term cardiotoxic side effects can develop in ~36% of patients up to 10 years after treatment. Despite being the most effective class of anti-cancer drug and widely used since last five decades, the molecular mechanisms that underly DIC remain poorly understood. To date, three major inter- related mechanisms for cardiotoxic effects of doxorubicin have been proposed: (i) generation of reactive oxygen species (ROS) and subsequent membrane damage, (ii) inhibition of topoisomerase II-β (TOP2B) topoisomerase I mitochondrial (TOP1MT), and (iii) modulation of intracellular calcium release. However, as cardiotoxicity in DIC patients may not emerge for years or decades, a better understanding of the different mechanisms in DIC across different cardiac cell types and their crosstalk can have significant implications on the search for therapeutics. The endothelium is a critical component of the cardiovascular system that forms a protective barrier for CMs and releases paracrine factors to maintain CM health and function. It has been shown that DOX disrupts the normal endothelial physiology by damaging ECs that can lead to the development of severe chronic vascular diseases such as atherosclerosis, which often leads to cardiac dysfunction. With the knowledge that dysfunctional ECs can have a negative impact on CM function, we need a better understanding of the integral role of ECs in the development of doxorubicin-induced myocardial injury. Despite impressive progress, little attention has been given to the potential importance of cell-to-cell signaling between ECs and CMs, despite the fact that ECs serve a paracrine function to enhance signaling in CMs, especially in context to pharmacological stimulation. This knowledge gap impedes our comprehensive understanding of organ dysfunction at a multi- cellular level. The overarching goal of our proposal is to use a multidisciplinary approach that integrates human iPSCs, bioengineering tools, and NGS to gain novel insights into the pathogenesis of DIC. We will pursue three specific aims. In Aim 1: we will establish an experimental platform to study the role of ECs in DIC. For this, we will recapitulate the EC-CM crosstalk in DIC patient’s iPSC-derived cells with 3D engineered heart tissues (EHTs). In Aim 2: we will decipher the mechanism of EC-CM crosstalk in EHTs treated with DOX using single- cell approaches (scRNA-seq and scATAC-seq). In Aim 3: we will validate the key regulatory players of EC-CM crosstalk in an animal model of DIC. Our proposal is supported by compelling preliminary data from a multi- disciplinary team of investigators. We believe we are well positioned to achieve the project goals within five years.