Unraveling the role of endothelium in chemotherapy-induced cardiotoxicity

项目摘要/摘要 阿霉素是一种高效的化疗药物，通常用于治疗多种癌症，但其用途是 因心脏毒性而受到限制。心脏毒性可从无症状的左心室射血功能降低 分数到高度症状性心力衰竭(III至IV级)。阿霉素急性心脏毒性(DIC) 发生在约11%的患者中，长期心脏毒性副作用可在~36%的患者中发展到10% 治疗数年后。尽管它是最有效的抗癌药物，并自过去五年以来被广泛使用 几十年来，导致DIC的分子机制仍然知之甚少。到目前为止，有三个主要的国际 阿霉素心脏毒性作用的相关机制已被提出：(I)产生反应性 氧物种(ROS)和随后的膜损伤，(Ii)拓扑异构酶II-β(TOP2B)的抑制 拓扑异构酶I线粒体(TOP1MT)，以及(Iii)细胞内钙释放的调节。然而，由于 DIC患者的心脏毒性可能在数年或数十年内不会出现，更好地了解不同 跨不同类型心肌细胞的DIC机制及其串扰可能对 对治疗学的探索。 血管内皮细胞是心血管系统的重要组成部分，对CMS形成保护屏障 并释放旁分泌因子以维持CM的健康和功能。已有研究表明，DOX可以扰乱 通过损伤内皮细胞而导致严重慢性血管的正常内皮生理 动脉粥样硬化等疾病，通常会导致心脏功能障碍。在了解到 功能障碍的ECs会对CM功能产生负面影响，我们需要更好地理解积分 内皮细胞在阿霉素致心肌损伤中的作用。尽管取得了令人瞩目的进展，但几乎没有 人们已经注意到ECs和CMS之间的细胞间信号的潜在重要性，尽管 事实上，ECs在CMS中发挥旁分泌功能以增强信号传递，特别是在药理学背景下 刺激。这种知识鸿沟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一种多学科的方法，将人类 IPSCs、生物工程工具和NGS，以获得对DIC发病机制的新见解。我们将追求三个目标 明确的目标。在目标1：我们将建立一个实验平台来研究ECs在DIC中的作用。为此，我们 将用3D工程心脏组织重述DIC患者IPSC来源细胞中的EC-CM串扰 (EHTS)。在目标2：我们将破译DOX治疗的EHTS中EC-CM串扰的机制。 细胞方法(scRNA-seq和scatac-seq)。在目标3：我们将验证EC-CM的主要监管参与者 DIC动物模型中的相声。我们的建议得到了令人信服的初步数据的支持，这些数据来自多个 纪律调查组。我们相信我们有能力在五年内实现项目目标 好几年了。

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.