Investigating the influence of biomimetic cues in cardiovascular system formation

组织结构及其力学性质是心脏和血管发育和功能的重要调节剂。我的研究项目包括相互关联的项目，这些项目专注于建立组织结构和生物力学力量的相关性，以及新血管和干细胞衍生的心脏细胞的成熟。在目标1中，我们将开发一种功能性的“微血管芯片”，以研究基质成分的影响以及在考虑流动的动态平台中对血管通透性中内皮和血管周围细胞-细胞相互作用的调节。需要创新的方法来产生功能性微血管，并以准确和可控的方式再现其生理微环境。我们将通过使用脂肪来源的微血管来实现这一点，这些微血管可以自组装成血管系统，并具有形成功能网络所需的所有细胞类型。该平台将首次允许在存在流动的情况下在体外生成复杂的功能性微血管;这代表了对现有策略的重大进步，并解决了这些应用的人类细胞来源方面的重要差距。在目标2中，我们将通过对血管细胞（内皮细胞和平滑肌细胞）施加周期性拉伸来定义牵引力在形成特定血管动脉身份中的影响，并研究它是否会优选地导致动脉身份的规范。我们将评估特定动脉基因的表达，血管周围募集，细胞面积，增殖和细胞间相互作用所涉及的生长因子。我们研究的创新来自于研究循环拉伸作为动脉身份正确分配的成熟线索，代表了一种潜在的新概念进步。由于心脏的发育依赖于血管的形成，因此在目的3中，我们将研究血管的存在是否促进了干细胞来源的心脏细胞的成熟。我们将通过确定在干细胞衍生的心脏细胞中加入血管或内皮细胞是否会促进其成熟来验证这一假设。将通过评估分子特征、结构和细胞组织、功能特性和电生理学来确定细胞成熟。这些将以来自不同发育阶段的人心肌细胞为基准。这是一个具有极高潜在影响的领域，在培养中获得人类成人样心脏细胞将是世界上第一个。总的来说，这些研究将扩大我们对心血管成熟和功能机制的理解，并将指导我们在体外产生复杂的，功能性血管化的人类心脏组织（我们的长期目标）的努力。

Tissue architecture and its mechanical properties are important modulators of the development and function of the heart and blood vessels. My research program consists of inter-related projects that are focused on establishing the relevance of tissue architecture and biomechanical forces, in the maturation of new blood vessels and of heart cells-derived from stem cells. In Objective 1, we will develop a functional “microvasculature-on-a-chip” to study effects of matrix composition and the modulation of endothelial and perivascular cell-cell interactions in vascular permeability in a dynamic platform that accounts for flow. Innovative approaches are needed for generating functional microvasculatures and recapitulating their physiological microenvironments in an accurate and controllable fashion. We will achieve this by use of adipose-derived microvessels that can self-assemble into a vasculature and have all the cell types necessary to form a functional network. This platform will allow for the first time, the generation of complex, functional microvasculatures in vitro in the presence of flow; which represents a significant advancement to existing strategies and solves an important gap in terms of human cell source for these applications. In Objective 2 we will define the influence of traction forces in the formation of specific vessel arterial identity by applying cyclic stretch to vascular cells (endothelial and smooth muscle cells) and investigating if it would preferably lead to the specification of arterial identity. We will assess the expression of specific arterial genes, growth factors involved in perivascular recruitment, cell area, proliferation and cell-cell interaction. The innovation in our studies comes from investigating cyclic stretch as a maturation cue for the proper assignment of arterial identity and represent a potential novel conceptual advance. Since the development of the heart is dependent on the formation of blood vessels, in Objective 3, we will investigate if the presence of blood vessels improves the maturation of the heart cells derived from stem cells. We will test this hypothesis by determining whether the addition of blood vessels or endothelial cells to stem cell-derived heart cells will enhance their maturation. Cell maturation will be determined by assessing the molecular signature, structural and cellular organization, functional properties and electrophysiology. These will be benchmarked against human cardiomyocytes from different stages of development. This is an area of extremely high potential impact where obtaining human adult-like heart cells in culture would be a world first. Collectively, these studies will expand our understanding of the mechanisms of cardiovascular maturation and function and will guide our efforts towards generating complex, functional vascularized human heart tissues in vitro (our long-term goal).