Unraveling the role of endothelium in chemotherapy-induced cardiotoxicity

揭示内皮在化疗引起的心脏毒性中的作用

• 批准号: 10340657
• 负责人: Nazish Sayed
• 金额: $ 39.35万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10340657
• 关键词: 3-Dimensional; Acute; Address; Affect; Aftercare; Animal Model; Anthracycline; Antineoplastic Agents; Atherosclerosis; Attention; Biochemical; Biomedical Engineering; CRISPR/Cas technology; Calcium; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cardiovascular Physiology; Cardiovascular system; Cells; Chromatin; Chronic; Coculture Techniques; Collaborations; Communication; Complex; Computational Biology; Data; Development; Dose; Doxorubicin; Endothelial Cells; Endothelium; Environment; Exhibits; Exposure to; Functional disorder; Gene Expression; Generations; Genes; Genomic DNA; Goals; Health; Heart; Heart failure; Human; Impairment; Kinetics; Knowledge; Lead; Left Ventricular Ejection Fraction; Ligands; Malignant Neoplasms; Mediating; Membrane; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Mus; Muscle Cells; Myocardial dysfunction; Oncology; Organ; Paracrine Communication; Pathogenesis; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiology; Play; Positioning Attribute; Reactive Oxygen Species; Research Personnel; Role; Signal Transduction; Site; Therapeutic; Topoisomerase II; Trastuzumab; Type I DNA Topoisomerases; Vascular Diseases; Wing; Work; base; cardiac tissue engineering; cell injury; cell type; chemotherapy; endothelial stem cell; gene regulatory network; genome editing; heart cell; improved; in vivo; induced pluripotent stem cell; induced pluripotent stem cell technology; insight; interdisciplinary approach; mechanical properties; mortality; mouse model; multidisciplinary; myocardial injury; next generation sequencing; novel; paracrine; predictive modeling; promoter; receptor; response; side effect; single cell technology; single-cell RNA sequencing; stem cell biology; tool; transcription factor

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.

项目摘要/摘要 阿霉素是一种通常用于治疗多种癌症的高效化学疗法药物，但其使用是 由于心脏毒性而受到限制。心脏毒性的范围从左心室射血的不对称降低范围 症状性心力衰竭的分数（III级至IV）。急性阿霉素诱导的心脏毒性（DIC） 发生在约11％的患者中，长期心脏毒性副作用可能会在约36％的患者中发展出来 治疗后几年。尽管是最有效的抗癌药物，并且自从过去五个以来广泛使用 几十年来，DIC下的分子机制仍然很少理解。迄今为止，三个主要之间 已经提出了阿霉素的心脏毒性作用的相关机制：（i）产生反应性 氧（ROS）和随后的膜损伤，（II）抑制拓扑异构酶II-β（TOP2B） 拓扑异构酶I线粒体（TOP1MT）和（iii）调节细胞内钙释放。但是，如 DIC患者的心脏毒性可能不会出现数年或几十年，对不同的了解 跨不同心脏细胞类型的DIC机制及其串扰可能对 寻找治疗。 内皮是心血管系统的关键组成部分，它形成了CMS的受保护障碍 并释放旁分泌因素以维持CM健康和功能。已经表明，DOX破坏了 正常的内皮生理学通过损害EC，从而导致严重慢性血管的发展 诸如动​​脉粥样硬化之类的疾病通常会导致心脏功能障碍。知道 功能失调的EC可能会对CM功能产生负面影响，我们需要更好地了解积分 EC在阿霉素诱导的心肌损伤中的作用。尽管进步令人印象深刻，但很少 注意EC和CMS之间的细胞间信号传导的潜在重要性，dospite ECS具有旁分泌功能以增强CMS的信号传导的事实，尤其是在药物的背景下 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。我们提案的总体目标是使用一种融合人类的多学科方法 IPSC，生物工程工具和NGS，以获得对DIC发病机理的新见解。我们将追求三个 具体目标。在AIM 1中：我们将建立一个实验平台来研究EC在DIC中的作用。为此，我们 将在DIC患者的IPSC衍生细胞中概括带有3D工程心脏组织的EC-CM串扰 （EHTS）。在AIM 2中：我们将使用单次使用DOX处理的EHT中EC-CM串扰的机理 细胞进近（SCRNA-SEQ和SCATAC-SEQ）。在AIM 3中：我们将验证EC-CM的关键监管参与者 在DIC动物模型中串扰。我们的建议得到了通过从多个多数数据中引人入胜的初步数据的支持 调查人员的纪律团队。我们认为，我们在五个之内实现项目目标的好处 年。