Oxylipin Signaling in Congenital Heart Disease

项目总结 先天性心脏病(CHD)是人类最常见的先天性缺陷类型， 然而，在大多数情况下，导致冠心病的原因尚不清楚。脂氧合酶衍生的氧脂是一种 一类复杂且特征不佳的高度氧化的多不饱和脂肪酸衍生生物活性 具有关键生物活性的脂类。我们最近发现了氧磷脂信号与人类 心血管发育和冠心病。这一发现是至关重要的，不仅是因为 知识，但也因为氧化脂是强烈的饮食依赖。我们的初步数据显示 脂氧合酶及其氧合脂质产物显著调节大鼠心脏中胚层的形成 人类心脏分化模型，它们的缺失会导致小鼠CHD。我们的中心假设是 内源性未定性的氧脂是心源性中胚层规格的关键调节因子 以及通过与Wnt信号通路的串扰来实现心脏的形态发生。我们的目标是 深入了解心脏发育过程中的脂氧合酶和氧脂信号转导途径 确定它们在CHD发病机制中的贡献。我们的具体目标是：(1)确定 人类心源性中胚层中氧脂信号的分子和细胞机制。 (2)研究氧化脂质在人干细胞心脏细胞外基质组织中的作用。 衍生心脏器官模型。(3)评价氧磷脂信号在先天性心脏病中的作用 小鼠的缺陷。方法：我们将研究人类多能干细胞来源的氧磷脂功能。 心脏中胚层和在3D心脏器官模型中结合LC-MS脂质组学，细胞和 分子生物学和生化方法。我们还将使用脂氧合酶基因敲除小鼠和 限制氧脂饮食以研究氧脂在小鼠胚胎体内心脏发育中的作用。 意义：这项工作通过探索细胞因子的生物学作用，解决了知识中的一个主要缺口 氧脂在心脏发育中的作用，以及新发现的与冠心病相关的机制。我们 也提供了饮食和先天性心脏病之间的新的潜在联系，具有重要的意义 用于预防冠心病。最后，我们提供了发现新的治疗方法的潜力，通过识别 可下药的目标和信号网络。创新：我们将首次探索关键 脂氧合酶、氧合脂与心脏发育信号网络的分子机制 人和老鼠的组织。我们还将建立一个新的心脏发育模式， 这些环境脂质在发育分子网络中的作用。影响：该项目将 极大地扩展心脏发育中的氧脂生物学知识，帮助制定策略 预防和潜在地治疗CHD，并为人类生物学中的氧脂研究开发新的工具。

Project summary Congenital heart defects (CHD) constitute the most common type of congenital defect in humans, however the causes for CHD in most cases are unknown. Lipoxygenase-derived oxylipins are a complex and poorly characterized class of highly oxidized, polyunsaturated fatty acid-derived bioactive lipids with key biological activities. We recently found a link between oxylipin signaling, human cardiovascular development and CHD. This finding is critical not only because of the existing gap in knowledge, but also because oxylipins are strongly dietary-dependent. Our preliminary data suggests lipoxygenases and their oxylipin products dramatically modulate cardiogenic mesoderm formation in human cardiac differentiation model, and their loss leads to CHD in mice. Our central hypothesis is that endogenous uncharacterized oxylipins are critical regulators of cardiogenic mesoderm specification and heart morphogenesis through cross-talk with the Wnt signaling pathway. Our objective is to provide insight into lipoxygenase and oxylipin signaling pathways during cardiac development and determine their contribution to mechanisms of CHD. Our Specific aims are: (1) Determine the molecular and cellular mechanisms of oxylipin signaling in human cardiogenic mesoderm specification. (2) Investigate the role of oxylipins in cardiac extracellular matrix organization in a human stem cell- derived cardiac organoid model. (3) Evaluate the contribution of oxylipin signaling to congenital heart defects in mice. Approach: We will study oxylipin function in human pluripotent stem cell-derived cardiogenic mesoderm and in a 3D cardiac organoid model by combining LC-MS lipidomics, cell and molecular biology and biochemical methods. We will also employ lipoxygenase knockout mice and oxylipin dietary restriction to study oxylipins in cardiac development in vivo in mouse embryos. Significance: This work addresses a major gap in knowledge by exploring the biological roles of oxylipins in cardiac development, and mechanisms underlying their newly discovered links to CHD. We also offer a novel potential link between diet and congenital heart defects with significant implications for CHD prevention. Finally, we offer the potential to discover new therapies for treatment by identifying druggable targets and signaling networks. Innovation: We will explore for the first time the key molecular mechanisms linking lipoxygenases, oxylipins and cardiac development signaling networks in human and mouse tissues. We will also establish a new model of cardiac development that integrates the role of these environmental lipids with developmental molecular networks. Impact: This project will dramatically expand knowledge of oxylipin biology in cardiac development, help develop strategies to prevent and potentially treat CHD, and develop new tools for the study of oxylipins in human biology.