Alcohol-induced cardiac injury and repair in human induced pluripotent stem cell model

项目摘要 酒精暴露是心脏发育异常的一个关键风险因素，并导致先天性心脏病， 婴儿死亡的主要非传染性原因。酒精诱导的心脏生长发育受损 早年生活也会增加成年后患心脏病的风险。此外，成人心脏系统也 对酒精敏感，这是一个重要但未被充分认识的危险因素， 疾病酒精暴露与缺血性事件、心律失常和心功能改变有关 和结构。这些病理后果可能是由酒精的复杂作用引起的， 增加的细胞毒性、氧化应激和异常的Ca 2+处理。酒精产生的炎性细胞因子 接触酒精也可能导致酒精引起的心脏损伤。然而，酒精的潜在机制- 诱发的心脏病尚未完全确定，缺乏有效的治疗方法。 传统上，由于可用性有限，关于酒精暴露的研究依赖于动物模型和细胞 和人原代心肌细胞的生长能力。然而，在动物模型中的研究是耗时的， 昂贵，并且不适合高通量药物筛选，并且由于生理学上的限制， 与人类的差异。为了补充对动物模型和细胞的研究，我们最近探索了 使用来源于人诱导多能干细胞（hiPSC-CM）的心肌细胞作为新的和 生理学相关模型，以研究酒精诱导的心脏毒性。hiPSC-CM具有许多类似于 人原代CM，可以被工程化为组织样结构并在长期培养中维持， 可以适用于高通量平台。 我们已经证明，暴露于临床相关剂量的酒精的hiPSC-CM可以概括 酒精暴露引起的病理事件，包括氧化应激、基因表达改变和心脏病 异常的Ca 2+处理和收缩性所指示的功能障碍。我们已经将hiPSC-CM改造成高- 通量格式和建立的高通量测定，以检测酒精诱导的心脏毒性和筛选 用于心脏增殖/保护剂。使用这种体外人类细胞模型和最先进的高- 通量技术，我们建议研究酒精诱导的毒性的潜在分子机制 并评估减轻酒精诱导的细胞毒性的潜在疗法。此外，本发明还提供了一种方法， 我们计划研究炎症细胞因子在酒精诱导的心脏损伤和修复中的作用。 我们期望我们建立的hiPSC-CM模型将有助于理解hiPSC-CM的分子和细胞机制 潜在的酒精毒性在人类心肌细胞和加速发展的针对性和有效的 治疗

PROJECT SUMMARY Alcohol exposure is a key risk factor for abnormal heart development and contributes to congenital heart disease, the leading non-infectious cause of infant mortality. Alcohol-induced impaired heart growth and development in early life can also increase the risk of heart disease later in adulthood. Furthermore, adult cardiac system is also sensitive to alcohol exposure, which is an important but underappreciated risk factor contributing to heart disease. Alcohol exposure is associated with ischemic events, arrhythmias and alterations in cardiac function and structure. These pathological consequences could result from complex actions of alcohol including increased cytotoxicity, oxidative stress and abnormal Ca2+ handling. Inflammatory cytokines produced by alcohol exposure could also contribute to alcohol-induced heart injury. However, mechanisms underlying alcohol- induced heart disease are not fully defined and effective therapies are lacking. Traditionally, studies on alcohol exposure have relied on animal models and cells because of limited availability and growth capacity of human primary cardiomyocytes. However, studies in animal models are time consuming, expensive, and not amenable for high-throughput drug screening, and have limitations due to physiological differences from humans. To complement the studies on animal models and cells, we have recently explored the use of cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) as a novel and physiologically relevant model to study alcohol-induced cardiotoxicity. hiPSC-CMs have many features similar to human primary CMs, can be engineered into tissue-like structures and maintained in long-term cultures, and can be adapted for high-throughput platforms. We have demonstrated that exposure of hiPSC-CMs with clinically relevant doses of alcohol can recapitulate pathological events caused by alcohol exposure, including oxidative stress, altered gene expression and cardiac dysfunction as indicated by abnormal Ca2+ handling and contractility. We have adapted hiPSC-CMs into high- throughput formats and established high-throughput assays to detect alcohol-induced cardiotoxicity and screen for cardiac proliferative/protective agents. Using this in vitro human cell model and state-of-the-art high- throughput technologies, we propose to investigate underlying molecular mechanisms of alcohol-induced toxicity in human cardiomyocytes and evaluate potential therapies to mitigate alcohol-induced cytotoxicity. In addition, we plan to investigate the involvement of inflammatory cytokines in alcohol-induced cardiac injury and repair. We expect that our established hiPSC-CM model will help understand the molecular and cellular mechanisms underlying alcohol toxicity in human cardiomyocytes and accelerate the development of targeted and effective therapies.